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HASUNUMA TomohisaEngineering Biology Research CenterProfessor
Researcher basic information
■ Research news- 06 May 2021, Microalgae biofuels: Changing carbohydrates into lipids
- 10 Mar. 2020, Successful method yielding high rate of D-lactate using cyanobacteria could revolutionize bioplastic production
- 17 Jan. 2020, Novel protein positioning technique improves functionality of yeast cells -Expected improvements in cell surface engineering across a variety of fields-
- 13 Dec. 2019, Success in metabolically engineering marine algae to synthesize valuable antioxidant astaxanthin
- 06 Sep. 2019, Fe metabolic engineering method succeeds in producing 1,2,4-butanetriol sustainably from biomass
- 06 Jun. 2018, World’s most efficient production of succinate from carbon dioxide
- 17 Apr. 2017, Making oil from algae ? towards more efficient biofuels
Research activity information
■ Award- Oct. 2022 神戸大学, 令和4年度 学長表彰(財務貢献者) 神戸大学
- Oct. 2021 神戸大学, 令和3年度 学長表彰(財務貢献者)
- Oct. 2020 神戸大学, 令和2年度 学長表彰(財務貢献者)
- Oct. 2019 神戸大学, 第11回学長表彰(財務貢献者)
- Oct. 2018 神戸大学, 第10回学長表彰(財務貢献者)
- Oct. 2017 神戸大学, 第9回学長表彰(財務貢献者)
- Sep. 2014 The Society for Biotechnology, Japan, Biotechnology Encouragement Prize, 代謝プロファイリングに基づく微生物育種技術の開発と応用
- Oct. 2013 Japan Bioindustry Association, Fermentation and Metabolism Research Prize, システムバイオロジー解析に基づく高機能型酵母創製とセルロースエタノール高生産技術開発への応用
- Springer Science and Business Media LLC, Jun. 2023, Archives of Microbiology, 205(7) (7), English[Refereed]Scientific journal
- Abstract l-Lactate is a major waste compound in cultured animal cells. To develop a sustainable animal cell culture system, we aimed to study the consumption of l-lactate using a photosynthetic microorganism. As genes involved in l-lactate utilization were not found in most cyanobacteria and microalgae, we introduced the NAD-independent l-lactate dehydrogenase gene from Escherichia coli (lldD) into Synechococcus sp. PCC 7002. The lldD-expressing strain consumed l-lactate added to basal medium. This consumption was accelerated by expression of a lactate permease gene from E. coli (lldP) and an increase in culture temperature. Intracellular levels of acetyl-CoA, citrate, 2-oxoglutarate, succinate, and malate, and extracellular levels of 2-oxoglutarate, succinate, and malate, increased during l-lactate utilization, suggesting that the metabolic flux from l-lactate was distributed toward the tricarboxylic acid cycle. This study provides a perspective on l-lactate treatment by photosynthetic microorganisms, which would increase the feasibility of animal cell culture industries.Springer Science and Business Media LLC, May 2023, Scientific Reports, 13(1) (1), English[Refereed]Scientific journal
- Elsevier BV, May 2023, Algal Research, 72, 103144 - 103144, English[Refereed]Scientific journal
- Wiley, Feb. 2023, Biotechnology and Bioengineering, 120(2) (2), 511 - 523, English[Refereed]Scientific journal
- Wiley, Jan. 2023, Biotechnology and Bioengineering, 120(4) (4), 1097 - 1107, English[Refereed]Scientific journal
- Abstract The bottleneck for the production of biofuels from microalgae consists on costly harvesting processes and low lipid production, immobilization technology could play a part on making the production of biofuels more feasible. The aim of this study was to evaluate the effect of alginate immobilization on the growth and lipid productivity of the microalgae Chlorella sorokiniana, so far, the main focus of immobilization technology has been its use for wastewater treatment and nutrient removal from effluents. The microalgae Chlorella sorokiniana was cultured in both free and immobilized forms under optimal autotrophic growth conditions. Microalgae were immobilized in calcium alginate beads generated by mixing algal cells with a sodium alginate solution, followed by extrusion into a CaCl2 solution. The results obtained in this study showed that the growth of the microalgae immobilized in alginate beads, was enhanced and achieved a dry cell weight 1.4-fold higher than that of a free cell culture, a higher light transmittance was also achieved in the alginate immobilized culture, and the lipid productivity was increased from 54.21 ± 2.48 mg l−1 d in the free cell culture to 82.22 ± 8.48 mg l−1 d in the immobilized culture. These results demonstrate the effectiveness of immobilization technology for promoting growth and lipid productivity in the microalgae Chlorella sorokiniana.IOP Publishing, Jan. 2023, Journal of Physics: Energy, 5(1) (1), 014019 - 014019, English[Refereed]Scientific journal
- Elsevier BV, Jan. 2023, Journal of Chromatography B, 1215, 123588 - 123588, English[Refereed]Scientific journal
- Synthetic methylotrophic Yarrowia lipolytica was constructed to convert methanol into biomass components and succinic acid.Royal Society of Chemistry (RSC), 2023, Green Chemistry, 25(1) (1), 183 - 195, English[Refereed]Scientific journal
- Abstract Photosynthesis must maintain stability and robustness throughout fluctuating natural environments. In cyanobacteria, dark-to-light transition leads to drastic metabolic changes from dark respiratory metabolism to CO2 fixation through the Calvin–Benson–Bassham (CBB) cycle using energy and redox equivalents provided by photosynthetic electron transfer. Previous studies have shown that catabolic metabolism supports the smooth transition into CBB cycle metabolism. However, metabolic mechanisms for robust initiation of photosynthesis are poorly understood due to lack of dynamic metabolic characterizations of dark-to-light transitions. Here, we show rapid dynamic changes (on a time scale of seconds) in absolute metabolite concentrations and 13C tracer incorporation after strong or weak light irradiation in the cyanobacterium Synechocystis sp. PCC 6803. Integration of this data enabled estimation of time-resolved nonstationary metabolic flux underlying CBB cycle activation. This dynamic metabolic analysis indicated that downstream glycolytic intermediates, including phosphoglycerate and phosphoenolpyruvate, accumulate under dark conditions as major substrates for initial CO2 fixation. Compared with wild-type Synechocystis, significant decreases in the initial oxygen evolution rate were observed in 12 h dark preincubated mutants deficient in glycogen degradation or oxidative pentose phosphate pathways. Accordingly, the degree of decrease in the initial oxygen evolution rate was proportional to the accumulated pool size of glycolytic intermediates. These observations indicate that the accumulation of glycolytic intermediates is essential for efficient metabolism switching under fluctuating light environments.Oxford University Press (OUP), Dec. 2022, Plant Physiology, English[Refereed]Scientific journal
- The four-carbon (C4) dicarboxylic acids, fumarate, malate, and succinate, are the most valuable targets that must be exploited for CO2-based chemical production in the move to a sustainable low-carbon future. Cyanobacteria excrete high amounts of C4 dicarboxylic acids through glycogen fermentation in a dark anoxic environment. The enhancement of metabolic flux in the reductive TCA branch in the Cyanobacterium Synechocystis sp. PCC6803 is a key issue in the C4 dicarboxylic acid production. To improve metabolic flux through the anaplerotic pathway, we have created the recombinant strain PCCK, which expresses foreign ATP-forming phosphoenolpyruvate carboxykinase (PEPck) concurrent with intrinsic phosphoenolpyruvate carboxylase (Ppc) overexpression. Expression of PEPck concurrent with Ppc led to an increase in C4 dicarboxylic acids by autofermentation. Metabolome analysis revealed that PEPck contributed to an increase in carbon flux from hexose and pentose phosphates into the TCA reductive branch. To enhance the metabolic flux in the reductive TCA branch, we examined the effect of corn-steep liquor (CSL) as a nutritional supplement on C4 dicarboxylic acid production. Surprisingly, the addition of sterilized CSL enhanced the malate production in the PCCK strain. Thereafter, the malate and fumarate excreted by the PCCK strain are converted into succinate by the CSL-settling microorganisms. Finally, high-density cultivation of cells lacking the acetate kinase gene showed the highest production of malate and fumarate (3.2 and 2.4 g/L with sterilized CSL) and succinate (5.7 g/L with non-sterile CSL) after 72 h cultivation. The present microbial community engineering is useful for succinate production by one-pot fermentation under dark anoxic conditions.American Chemical Society (ACS), Dec. 2022, ACS Synthetic Biology, 11(12) (12), 4054 - 4064, English, International magazine[Refereed]Scientific journal
- Elsevier BV, Dec. 2022, Biochemical Engineering Journal, 188, 108681 - 108681, English[Refereed]Scientific journal
- Abstract Engineering the microbial production of secondary metabolites is limited by the known reactions of correctly annotated enzymes. Therefore, the machine learning discovery of specialized enzymes offers great potential to expand the range of biosynthesis pathways. Benzylisoquinoline alkaloid production is a model example of metabolic engineering with potential to revolutionize the paradigm of sustainable biomanufacturing. Existing bacterial studies utilize a norlaudanosoline pathway, whereas plants contain a more stable norcoclaurine pathway, which is exploited in yeast. However, committed aromatic precursors are still produced using microbial enzymes that remain elusive in plants, and additional downstream missing links remain hidden within highly duplicated plant gene families. In the current study, machine learning is applied to predict and select plant missing link enzymes from homologous candidate sequences. Metabolomics-based characterization of the selected sequences reveals potential aromatic acetaldehyde synthases and phenylpyruvate decarboxylases in reconstructed plant gene-only benzylisoquinoline alkaloid pathways from tyrosine. Synergistic application of the aryl acetaldehyde producing enzymes results in enhanced benzylisoquinoline alkaloid production through hybrid norcoclaurine and norlaudanosoline pathways.Springer Science and Business Media LLC, Dec. 2022, Nature Communications, 13(1) (1), 1405, EnglishScientific journal
- Abstract Background Microalgal lipid production has attracted global attention in next-generation biofuel research. Nitrogen starvation, which drastically suppresses cell growth, is a common and strong trigger for lipid accumulation in microalgae. We previously developed a mutant Chlamydomonas sp. KAC1801, which can accumulate lipids irrespective of the presence or absence of nitrates. This study aimed to develop a feasible strategy for stable and continuous lipid production through semi-continuous culture of KAC1801. Results KAC1801 continuously accumulated > 20% lipid throughout the subculture (five generations) when inoculated with a dry cell weight of 0.8–0.9 g L−1 and cultured in a medium containing 18.7 mM nitrate, whereas the parent strain KOR1 accumulated only 9% lipid. Under these conditions, KAC1801 continuously produced biomass and consumed nitrates. Lipid productivity of 116.9 mg L−1 day−1 was achieved by semi-continuous cultivation of KAC1801, which was 2.3-fold higher than that of KOR1 (50.5 mg L−1 day−1). Metabolome and transcriptome analyses revealed a depression in photosynthesis and activation of nitrogen assimilation in KAC1801, which are the typical phenotypes of microalgae under nitrogen starvation. Conclusions By optimizing nitrate supply and cell density, a one-step cultivation system for Chlamydomonas sp. KAC1801 under nitrate-replete conditions was successfully developed. KAC1801 achieved a lipid productivity comparable to previously reported levels under nitrogen-limiting conditions. In the culture system of this study, metabolome and transcriptome analyses revealed a nitrogen starvation-like response in KAC1801.Springer Science and Business Media LLC, Sep. 2022, Biotechnology for Biofuels and Bioproducts, 15(1) (1), English[Refereed]Scientific journal
- Springer Science and Business Media LLC, Sep. 2022, Applied Microbiology and Biotechnology, 106(18) (18), 6347 - 6361, English[Refereed]Scientific journal
- Wiley, Sep. 2022, Microbial Biotechnology, 15(9) (9), 2364 - 2378, English[Refereed]Scientific journal
- BACKGROUND: Glutathione is a valuable tri-peptide that is industrially produced by fermentation using the yeast Saccharomyces cerevisiae, and is widely used in the pharmaceutical, food, and cosmetic industries. It has been reported that addition of L-serine (L-Ser) is effective at increasing the intracellular glutathione content because L-Ser is the common precursor of L-cysteine (L-Cys) and glycine (Gly) which are substrates for glutathione biosynthesis. Therefore, we tried to enhance the L-Ser biosynthetic pathway in S. cerevisiae for improved glutathione production. RESULTS: The volumetric glutathione production of recombinant strains individually overexpressing SER2, SER1, SER3, and SER33 involved in L-Ser biosynthesis at 48 h cultivation was increased 1.3, 1.4, 1.9, and 1.9-fold, respectively, compared with that of the host GCI strain, which overexpresses genes involved in glutathione biosynthesis. We further examined simultaneous overexpression of SHM2 and/or CYS4 genes involved in Gly and L-Cys biosynthesis, respectively, using recombinant GCI strain overexpressing SER3 and SER33 as hosts. As a result, GCI overexpressing SER3, SHM2, and CYS4 showed the highest volumetric glutathione production (64.0 ± 4.9 mg/L) at 48 h cultivation, and this value is about 2.5-fold higher than that of the control strain. CONCLUSIONS: This study first revealed that engineering of L-Ser and Gly biosynthetic pathway are useful strategies for fermentative glutathione production by S. cerevisiase.Springer Science and Business Media LLC, Aug. 2022, Microbial cell factories, 21(1) (1), 153 - 153, English, International magazineScientific journal
- American Chemical Society (ACS), Jun. 2022, ACS Synthetic Biology, 11(6) (6), 2098 - 2107, EnglishScientific journal
- Abstract Expression of secreted recombinant proteins burdens the protein secretion machinery, limiting production. Here, we describe an approach to improving protein production by the non-conventional yeast Komagataella phaffii comprised of genome-wide screening for effective gene disruptions, combining them in a single strain, and recovering growth reduction by adaptive evolution. For the screen, we designed a multiwell-formatted, streamlined workflow to high-throughput assay of secretion of a single-chain small antibody, which is cumbersome to detect but serves as a good model of proteins that are difficult to secrete. Using the consolidated screening system, we evaluated >19,000 mutant strains from a mutant library prepared by a modified random gene-disruption method, and identified six factors for which disruption led to increased antibody production. We then combined the disruptions, up to quadruple gene knockouts, which appeared to contribute independently, in a single strain and observed an additive effect. Target protein and promoter were basically interchangeable for the effects of knockout genes screened. We finally used adaptive evolution to recover reduced cell growth by multiple gene knockouts and examine the possibility for further enhancing protein secretion. Our successful, three-part approach holds promise as a method for improving protein production by non-conventional microorganisms.Springer Science and Business Media LLC, Jun. 2022, Communications Biology, 5(1) (1), 561, English[Refereed]Scientific journal
- Informa UK Limited, Apr. 2022, Journal of Inflammation Research, Volume 15, 2387 - 2395, English[Refereed]Scientific journal
- Elsevier BV, Feb. 2022, Current Opinion in Green and Sustainable Chemistry, 33, 100584 - 100584, EnglishScientific journal
- Elsevier BV, Jan. 2022, Biochemical Engineering Journal, 178, 108296 - 108296Scientific journal
- Elsevier BV, Jan. 2022, Biochemical Engineering Journal, 178, 108274 - 108274Scientific journal
- Elsevier BV, Jan. 2022, Biotechnology Notes, 3, 1 - 7, EnglishScientific journal
- Elsevier BV, Jan. 2022, Bioresource Technology, 344, 126196 - 126196, EnglishScientific journal
- Elsevier BV, Jan. 2022, Bioresource Technology, 343, 126071 - 126071, EnglishScientific journal
- Nitrogen is essential for the biosynthesis of various molecules in cells, such as amino acids and nucleotides, as well as several types of lipids and sugars. Cyanobacteria can assimilate several forms of nitrogen, including nitrate, ammonium, and urea, and the physiological and genetic responses to these nitrogen sources have been studied previously. However, the metabolic changes in cyanobacteria caused by different nitrogen sources have not yet been characterized. This study aimed to elucidate the influence of nitrate and ammonium on the metabolic profiles of the cyanobacterium Synechocystis sp. strain PCC 6803. When supplemented with NaNO3 or NH4Cl as the nitrogen source, Synechocystis sp. PCC 6803 grew faster in NH4Cl medium than in NaNO3 medium. Metabolome analysis indicated that some metabolites in the CBB cycle, glycolysis, and TCA cycle, and amino acids were more abundant when grown in NH4Cl medium than NaNO3 medium. 15N turnover rate analysis revealed that the nitrogen assimilation rate in NH4Cl medium was higher than in NaNO3 medium. These results indicate that the mechanism of nitrogen assimilation in the GS-GOGAT cycle differs between NaNO3 and NH4Cl. We conclude that the amounts and biosynthetic rate of cyanobacterial metabolites varies depending on the type of nitrogen.MDPI AG, Dec. 2021, Metabolites, 11(12) (12), 867 - 867Scientific journal
- Springer Science and Business Media LLC, Dec. 2021, Microbial Cell Factories, 20(1) (1), 228, English
Abstract Background Bio-based aromatic compounds are of great interest to the industry, as commercial production of aromatic compounds depends exclusively on the unsustainable use of fossil resources or extraction from plant resources. γ-amino acid 3-amino-4-hydroxybenzoic acid (3,4-AHBA) serves as a precursor for thermostable bioplastics.Results Under aerobic conditions, a recombinantCorynebacterium glutamicum strain KT01 expressinggriH andgriI genes derived fromStreptomyces griseus produced 3,4-AHBA with large amounts of amino acids as by-products. The specific productivity of 3,4-AHBA increased with decreasing levels of dissolved oxygen (DO) and was eightfold higher under oxygen limitation (DO = 0 ppm) than under aerobic conditions (DO ≥ 2.6 ppm). Metabolic profiles during 3,4-AHBA production were compared at three different DO levels (0, 2.6, and 5.3 ppm) using the DO-stat method. Results of the metabolome analysis revealed metabolic shifts in both the central metabolic pathway and amino acid metabolism at a DO of < 33% saturated oxygen. Based on this metabolome analysis, metabolic pathways were rationally designed for oxygen limitation. Anldh deletion mutant, with the loss of lactate dehydrogenase, exhibited 3.7-fold higher specific productivity of 3,4-AHBA at DO = 0 ppm as compared to the parent strain KT01 and produced 5.6 g/L 3,4-AHBA in a glucose fed-batch culture.Conclusions Our results revealed changes in the metabolic state in response to DO concentration and provided insights into oxygen supply during fermentation and the rational design of metabolic pathways for improved production of related amino acids and their derivatives.Graphical Abstract Scientific journal - Elsevier BV, Dec. 2021, Algal Research, 60, 102544 - 102544Scientific journal
- Elsevier BV, Dec. 2021, Metabolic Engineering Communications, 13, e00188 - e00188, EnglishScientific journal
- Springer Science and Business Media LLC, Dec. 2021, Biotechnology for Biofuels, 14(1) (1), 39 - 39, English, International magazine
Abstract Background Trichoderma reesei is a filamentous fungus that is important as an industrial producer of cellulases and hemicellulases due to its high secretion of these enzymes and outstanding performance in industrial fermenters. However, the reduction of enzyme production caused by carbon catabolite repression (CCR) has long been a problem. Disruption of a typical transcriptional regulator, Cre1, does not sufficiently suppress this reduction in the presence of glucose.Results We found that deletion of an α-tubulin (tubB ) inT. reesei enhanced both the amount and rate of secretory protein production. Also, the tubulin-disrupted (ΔtubB) strain had high enzyme production and the same enzyme profile even if the strain was cultured in a glucose-containing medium. From transcriptome analysis, the ΔtubB strain exhibited upregulation of both cellulase and hemicellulase genes including some that were not originally induced by cellulose. Moreover, cellobiose transporter genes and the other sugar transporter genes were highly upregulated, and simultaneous uptake of glucose and cellobiose was also observed in the ΔtubB strain. These results suggested that the ΔtubB strain was released from CCR.Conclusion Trichoderma reesei α-tubulin is involved in the transcription of cellulase and hemicellulase genes, as well as in CCR. This is the first report of overcoming CCR by disrupting α-tubulin gene inT. reesei . The disruption of α-tubulin is a promising approach for creating next-generation enzyme-producing strains ofT. reesei. Scientific journal - Springer Science and Business Media LLC, Dec. 2021, Communications Biology, 4(1) (1)
Abstract Light/dark cycling is an inherent condition of outdoor microalgae cultivation, but is often unfavorable for lipid accumulation. This study aims to identify promising targets for metabolic engineering of improved lipid accumulation under outdoor conditions. Consequently, the lipid-rich mutantChlamydomonas sp. KOR1 was developed through light/dark-conditioned screening. During dark periods with depressed CO2 fixation, KOR1 shows rapid carbohydrate degradation together with increased lipid and carotenoid contents. KOR1 was subsequently characterized with extensive mutation of theISA1 gene encoding a starch debranching enzyme (DBE). Dynamic time-course profiling and metabolomics reveal dramatic changes in KOR1 metabolism throughout light/dark cycles. During light periods, increased flux from CO2 through glycolytic intermediates is directly observed to accompany enhanced formation of small starch-like particles, which are then efficiently repartitioned in the next dark cycle. This study demonstrates that disruption of DBE can improve biofuel production under light/dark conditions, through accelerated carbohydrate repartitioning into lipid and carotenoid.Scientific journal - Elsevier BV, Nov. 2021, Bioresource Technology, 340, 125638 - 125638, EnglishScientific journal
- Elsevier BV, Sep. 2021, Energy Conversion and Management, 243, 114359 - 114359, EnglishScientific journal
- Springer Science and Business Media LLC, Aug. 2021, Applied Microbiology and Biotechnology, 105(14-15) (14-15), 5895 - 5904, EnglishScientific journal
- Elsevier BV, Aug. 2021, Bioresource Technology, 334, 125200 - 125200, EnglishScientific journal
- In the past few decades, microalgae-based bioremediation methods for treating heavy metal (HM)-polluted wastewater have attracted much attention by virtue of their environment friendliness, cost efficiency, and sustainability. However, their HM removal efficiency is far from practical use. Directed evolution is expected to be effective for developing microalgae with a much higher HM removal efficiency, but there is no non-invasive or label-free indicator to identify them. Here, we present an intelligent cellular morphological indicator for identifying the HM removal efficiency of Euglena gracilis in a non-invasive and label-free manner. Specifically, we show a strong monotonic correlation (Spearman's ρ = -0.82, P = 2.1 × 10-5) between a morphological meta-feature recognized via our machine learning algorithms and the Cu2+ removal efficiency of 19 E. gracilis clones. Our findings firmly suggest that the morphology of E. gracilis cells can serve as an effective HM removal efficiency indicator and hence have great potential, when combined with a high-throughput image-activated cell sorter, for directed-evolution-based development of E. gracilis with an extremely high HM removal efficiency for practical wastewater treatment worldwide.American Chemical Society (ACS), Jun. 2021, Environmental Science & Technology, 55(12) (12), 7880 - 7889, English, International magazineScientific journal
- Reconstitution of prenylflavonoids using the flavonoid biosynthetic pathway and prenyltransferases (PTs) in microbes can be a promising attractive alternative to plant-based production or chemical synthesis. Here, we demonstrate that promiscuous microbial PTs can be a substitute for regiospecific but mostly unidentified botanical PTs. To test the prenylations of naringenin, we constructed a yeast strain capable of producing naringenin from l-phenylalanine by genomic integration of six exogenous genes encoding components of the naringenin biosynthetic pathway. Using this platform strain, various microbial PTs were tested for prenylnaringenin production. In vitro screening demonstrated that the fungal AnaPT (a member of the tryptophan dimethylallyltransferase family) specifically catalyzed C-3' prenylation of naringenin, whereas SfN8DT-1, a botanical PT, specifically catalyzed C-8 prenylation. In vivo, the naringenin-producing strain expressing the microbial AnaPT exhibited heterologous microbial production of 3'-prenylnaringenin (3'-PN), in contrast to the previously reported in vivo production of 8-prenylnaringenin (8-PN) using the botanical SfN8DT-1. These findings provide strategies towards expanding the production of a variety of prenylated compounds, including well-known prenylnaringenins and novel prenylflavonoids. These results also suggest the opportunity for substituting botanical PTs, both known and unidentified, that display relatively strict regiospecificity of the prenyl group transfer.Elsevier BV, Jun. 2021, Metabolic Engineering Communications, 12, e00169 - e00169, English, International magazineScientific journal
- Elsevier BV, May 2021, Trends in Biotechnology, 39(5) (5), 488 - 504, EnglishScientific journal
- Oxford University Press (OUP), Apr. 2021, Bioscience, Biotechnology, and Biochemistry, 85(5) (5), 1275 - 1282, English
ABSTRACT Streptomyces incarnatus NRRL8089 produces the antiviral, antifungal, antiprotozoal nucleoside antibiotic sinefungin. To enhance sinefungin production, multiple mutations were introduced to the rpoB gene encoding RNA polymerase (RNAP) β-subunit at the target residues, D447, S453, H457, and R460. Sparse regression analysis using elastic-net lasso-ridge penalties on previously reported H457X mutations identified a numeric parameter set, which suggested that H457R/Y/F may cause production enhancement. H457R/R460C mutation successfully enhanced the sinefungin production by 3-fold, while other groups of mutations, such as D447G/R460C or D447G/H457Y, made moderate or even negative effects. To identify why the rif cluster residues have diverse effects on sinefungin production, an RNAP/DNA/mRNA complex model was constructed by homology modeling and molecular dynamics simulation. The 4 residues were located near the mRNA strand. Density functional theory–based calculation suggested that D447, H457, and R460 are in direct contact with ribonucleotide, and partially positive charges are induced by negatively charged chain of mRNA.Scientific journal - Wiley, Jan. 2021, Biotechnology and Bioengineering, 118(1) (1), 175 - 185, EnglishScientific journal
- Elsevier BV, Jan. 2021, Talanta, 222, 121625 - 121625, EnglishScientific journal
- Oxford University Press (OUP), Dec. 2020, Nucleic Acids Research, 48(22) (22), 13000 - 13012, English
Abstract In the yeast Saccharomyces cerevisiae, terminator sequences not only terminate transcription but also affect expression levels of the protein-encoded upstream of the terminator. The non-conventional yeast Pichia pastoris (syn. Komagataella phaffii) has frequently been used as a platform for metabolic engineering but knowledge regarding P. pastoris terminators is limited. To explore terminator sequences available to tune protein expression levels in P. pastoris, we created a ‘terminator catalog’ by testing 72 sequences, including terminators from S. cerevisiae or P. pastoris and synthetic terminators. Altogether, we found that the terminators have a tunable range of 17-fold. We also found that S. cerevisiae terminator sequences maintain function when transferred to P. pastoris. Successful tuning of protein expression levels was shown not only for the reporter gene used to define the catalog but also using betaxanthin production as an example application in pathway flux regulation. Moreover, we found experimental evidence that protein expression levels result from mRNA abundance and in silico evidence that levels reflect the stability of mRNA 3′-UTR secondary structure. In combination with promoter selection, the novel terminator catalog constitutes a basic toolbox for tuning protein expression levels in metabolic engineering and synthetic biology in P. pastoris.Scientific journal - Dec. 2020, Journal of Proteome Data and Methods, 2(0005) (0005), EnglishJapan Computational Mass Spectrometry Meeting 2020 Activity ReportScientific journal
- Springer Science and Business Media LLC, Dec. 2020, Nature Communications, 11(1) (1), 3452, English[Refereed]Scientific journal
- Cold Spring Harbor Laboratory, Jul. 2020, Biotechnology for Biofuels, 13(1) (1), 1 - 15, English
Abstract Consolidated bioprocessing, which combines saccharolytic and fermentative abilities in a single microorganism, is receiving increased attention to decrease environmental and economic costs in lignocellulosic biorefineries. Nevertheless, the economic viability of lignocellulosic ethanol is also dependent of an efficient utilization of the hemicellulosic fraction, which is mainly composed of xylose and may comprise up to 40 % of the total biomass. This major bottleneck is mainly due to the necessity of chemical/enzymatic treatments to hydrolyze hemicellulose into fermentable sugars and to the fact that xylose is not readily consumed bySaccharomyces cerevisiae – the most used organism for large-scale ethanol production. In this work, industrialS. cerevisiae strains, presenting robust traits such as thermotolerance and improved resistance to inhibitors, were evaluated as hosts for the cell-surface display of hemicellulolytic enzymes and optimized xylose assimilation, aiming at the development of whole-cell biocatalysts for consolidated bioprocessing of corn cob-derived hemicellulose. These modifications allowed the direct production of ethanol from non-detoxified hemicellulosic liquor obtained by hydrothermal pretreatment of corn cob, reaching an ethanol titer of 11.1 g/L corresponding to a yield of 0.328 gram per gram of potential xylose and glucose, without the need for external hydrolytic catalysts. Also, consolidated bioprocessing of pretreated corn cob was found to be more efficient for hemicellulosic ethanol production than simultaneous saccharification and fermentation with addition of commercial hemicellulases. These results show the potential of industrialS. cerevisiae strains for the design of whole-cell biocatalysts and paves the way for the development of more efficient consolidated bioprocesses for lignocellulosic biomass valorization, further decreasing environmental and economic costs.[Refereed]Scientific journal - Elsevier BV, Jun. 2020, Bioresource Technology, 305, 123072 - 123072, English[Refereed]Scientific journal
- May 2020, Science Advances, 6, eaba6712, EnglishSequentially addressable dielectrophoretic array for high-throughput sorting of large-volume biological compartments[Refereed]Scientific journal
- d-Lactate is one of the most valuable compounds for manufacturing biobased polymers. Here, we have investigated the significance of endogenous malate dehydrogenase (decarboxylating) (malic enzyme, ME), which catalyzes the oxidative decarboxylation of malate to pyruvate, in d-lactate biosynthesis in the cyanobacterium Synechocystis sp. PCC6803. d-Lactate levels were increased by 2-fold in ME-overexpressing strains, while levels in ME-deficient strains were almost equivalent to those in the host strain. Dynamic metabolomics revealed that overexpression of ME led to increased turnover rates in malate and pyruvate metabolism; in contrast, deletion of ME resulted in increased pool sizes of glycolytic intermediates, probably due to sequential feedback inhibition, initially triggered by malate accumulation. Finally, both the loss of the acetate kinase gene and overexpression of endogenous d-lactate dehydrogenase, concurrent with ME overexpression, resulted in the highest production of d-lactate (26.6 g/L) with an initial cell concentration of 75 g-DCW/L after 72 h fermentation.Feb. 2020, ACS synthetic biology, 9(2) (2), 260 - 268, English, International magazine[Refereed]
- Jan. 2020, Metabolic Engineering, 57, 110 - 117, EnglishNovel strategy for anchorage position control of GPI-attached proteins in the yeast cell wall using different GPI-anchoring domains[Refereed]Scientific journal
- Jan. 2020, Trends in Biotechnology, 38(1) (1), 68 - 82, EnglishDynamic metabolomics for engineering biology: Accelerating learning cycles for bioproduction[Refereed]Scientific journal
- Dec. 2019, Metabolic Engineering, 56, 17 - 27, EnglishProduction of 1,2,4-butanetriol from xylose by Saccharomyces cerevisiae through Fe metabolic engineering[Refereed]Scientific journal
- © 2019, The Author(s). Previous studies have utilized monoamine oxidase (MAO) and L-3,4-dihydroxyphenylalanine decarboxylase (DDC) for microbe-based production of tetrahydropapaveroline (THP), a benzylisoquinoline alkaloid (BIA) precursor to opioid analgesics. In the current study, a phylogenetically distinct Bombyx mori 3,4-dihydroxyphenylacetaldehyde synthase (DHPAAS) is identified to bypass MAO and DDC for direct production of 3,4-dihydroxyphenylacetaldehyde (DHPAA) from L-3,4-dihydroxyphenylalanine (L-DOPA). Structure-based enzyme engineering of DHPAAS results in bifunctional switching between aldehyde synthase and decarboxylase activities. Output of dopamine and DHPAA products is fine-tuned by engineered DHPAAS variants with Phe79Tyr, Tyr80Phe and Asn192His catalytic substitutions. Balance of dopamine and DHPAA products enables improved THP biosynthesis via a symmetrical pathway in Escherichia coli. Rationally engineered insect DHPAAS produces (R,S)-THP in a single enzyme system directly from L-DOPA both in vitro and in vivo, at higher yields than that of the wild-type enzyme. However, DHPAAS-mediated downstream BIA production requires further improvement.Dec. 2019, Nature Communications, 10(1) (1)[Refereed]Scientific journal
- Dec. 2019, Metabolites, 9(12) (12), 297, EnglishShort-term temporal metabolic behavior in halophilic cyanobacterium Synechococcus sp. strain PCC 7002 after salt shock[Refereed]Scientific journal
- Nov. 2019, Green Chemistry, 22(1) (1), 153 - 162, EnglishFermentation of pigment-extracted microalgal residue using yeast cell-surface display: direct high-density ethanol production with competitive life cycle impacts[Refereed]Scientific journal
- Oct. 2019, Bioresource Technology, 290, 121786, EnglishA novel process for the mixotrophic production of lutein with Chlorella sorokiniana MB-1-M12 using aquaculture wastewater[Refereed]Scientific journal
- Oct. 2019, ACS Synthetic Biology, 8, 2701 - 2709, EnglishSingle-Stage Astaxanthin Production Enhances the Nonmevalonate Pathway and Photosynthetic Central Metabolism in Synechococcus sp. PCC 7002[Refereed]Scientific journal
- Sep. 2019, Biotechnology Journal, 14(9) (9), e1800704, EnglishCombined cell surface display of β-D-glucosidase (BGL), maltose transporter (MAL11) and overexpression of cytosolic xylose reductase (XR) in Saccharomyces cerevisiae enhance cellobiose/xylose co-utilization for xylitol bio-production from lignocellulosic[Refereed]Scientific journal
- Jul. 2019, mBio, 10(4) (4), e00833-19, EnglishDay/night separation of oxygenic energy metabolism and nuclear DNA replication in the unicellular red alga Cyanidioschyzon merolae[Refereed]Scientific journal
- May 2019, Green Chemistry, 21, 1795 - 1808, EnglishCell-surface display technology and metabolic engineering of Saccharomyces cerevisiae for enhancing xylitol production from woody biomass[Refereed]Scientific journal
- May 2019, Nature Communications, 10, 2015, EnglishMechanism-based tuning of insect 3,4-dihydroxyphenylacetaldehyde synthase for synthetic bioproduction of benzylisoquinoline alkaloids[Refereed]Scientific journal
- Apr. 2019, Bioresource Technology, 278, 17 - 25, EnglishEnhancing lutein production with mixotrophic cultivation of Chlorella sorokiniana MB-1-M12 using different bioprocess operation strategies[Refereed]Scientific journal
- Yeasts are extremely useful, not only for fermentation but also for a wide spectrum of fuel and chemical productions. We analyzed the overall metabolic turnover and transcript dynamics in glycolysis and the TCA cycle, revealing the difference in adaptive pyruvate metabolic response between a Crabtree-negative species, Kluyveromyces marxianus, and a Crabtree-positive species, Saccharomyces cerevisiae, during aerobic growth. Pyruvate metabolism was inclined toward ethanol production under aerobic conditions in S. cerevisiae, while increased transcript abundances of the genes involved in ethanol metabolism and those encoding pyruvate dehydrogenase were seen in K. marxianus, indicating the augmentation of acetyl-CoA synthesis. Furthermore, different metabolic turnover in the TCA cycle was observed in the two species: malate and fumarate production in S. cerevisiae was higher than in K. marxianus, irrespective of aeration; however, fluxes of both the reductive and oxidative TCA cycles were enhanced in K. marxianus by aeration, implying both the cycles contribute to efficient electron flux without producing ethanol. Additionally, decreased hexokinase activity under aerobic conditions is expected to be important for maintenance of suitable carbon flux. These findings demonstrate differences in the key metabolic trait of yeasts employing respiration or fermentation, and provide important insight into the metabolic engineering of yeasts.Mar. 2019, Scientific Reports, 9(1) (1), 5319 - 5319, English, International magazine[Refereed]Scientific journal
- Feb. 2019, Applied Microbiology and Biotechnology, 103(3) (3), 1243 - 1254, EnglishSustainable production of glutathione from lignocellulose-derived sugars using engineered Saccharomyces cerevisiae[Refereed]Scientific journal
- Feb. 2019, Biotechnology for Biofuels, 12, 39, EnglishLight/dark cycling causes delayed lipid accumulation and increased photoperiod-based biomass yield by altering metabolic flux in oleaginous Chlamydomonas sp.[Refereed]Scientific journal
- Feb. 2019, Biochemical Engineering Journal, 142, 117 - 123, English5-Hydroxymethylfurfural production from salt-induced photoautotrophically cultivated Chlorella sorokiniana[Refereed]Scientific journal
- Oxford University Press (OUP), Nov. 2018, FEMS Yeast Research, 18(7) (7), foy074, English[Refereed]Scientific journal
- Sep. 2018, Microbial Cell Factories, 17, 153, EnglishWidespread effect of N-acetyl-D-glucosamine assimilation on the metabolisms of amino acids, purines, and pyrimidines in Scheffersomyces stipitis[Refereed]Scientific journal
- Aug. 2018, eLife, 7, e34595, EnglishEstimating the protein burden limit of yeast cells by measuring expression limits of glycolytic proteins[Refereed]Scientific journal
- Succinate is a versatile petrochemical compound that can be produced by microorganisms, often from carbohydrate based carbon sources. Phototrophic cyanobacteria including Synechocystis sp. PCC 6803 can more efficiently produce organic acids such as succinate without sugar supplementation, via photosynthetic production of glycogen followed by glycogen utilization, typically under dark conditions. In this study, Synechocystis 6803 bioproduction of organic acids under dark anoxic conditions was found to increase with elevation of temperature from 30 °C to 37 °C. The further enhancement of succinate bioproduction by overexpression of the rate limiting enzyme phosphoenolpyruvate carboxylase resulted in improved glycogen utilization. To gain more insight into the mechanisms underlying the increased organic acid output, a novel temperature dependent metabolomics analysis was performed. Adenylate energy charge was found to decrease along with elevating temperature, while central metabolites glucose 6-phosphate, fructose 6-phosphate, fructose 1,6-bisphosphate, glycerol 3-phosphate, malate, fumarate and succinate increased. Temperature dependent 13C-labeling metabolomics analysis further revealed a glycolysis to TCA bottleneck, which could be overcome by addition of CO2, leading to even higher organic acid production. Optimization of initial cell concentration to 25 g-dry cell weight/L, in combination with 100 mM NaHCO3 supplementation, afforded a succinate titer of over 1.8 g/L, the highest reported autotrophic succinate titer. Succinate titers remained high after additional knockout of ackA, resulting in the highest reported autotrophic D-lactate titer as well. The optimization of Synechocystis 6803 organic acid production therefore holds significant promise for CO2 capture and utilization.Academic Press Inc., Jul. 2018, Metabolic Engineering, 48, 109 - 120, English[Refereed]Scientific journal
- American Society for Microbiology, Jul. 2018, Applied and Environmental Microbiology, 84(15) (15), e02882 - 17, English
ABSTRACT The methylotrophic yeastPichia pastoris is widely used to produce recombinant proteins, taking advantage of this species' high-density cell growth and strong ability to secrete proteins. Circular plasmids containing theP. pastoris -specific autonomously replicating sequence (PARS1 ) permit transformation ofP. pastoris with higher efficiency than obtained following chromosomal integration by linearized DNA. Unfortunately, however, existing autonomously replicating plasmids are known to be inherently unstable. In this study, we used transcriptome sequencing (RNA-seq) data and genome sequence information to independently identify, on each of the four chromosomes, centromeric DNA sequences consisting of long inverted repeat sequences. By examining the chromosome 2 centromeric DNA sequence (Cen2 ) in detail, we demonstrate that an ∼111-bp region located at one end of the putative centromeric sequence had autonomous replication activity. In addition, the full-lengthCen2 sequence, which contains two long inverted repeat sequences and a nonrepetitive central core region, is needed for the accurate replication and distribution of plasmids inP. pastoris . Thus, we constructed a new, stable, autonomously replicating plasmid vector that harbors the entireCen2 sequence; this episome facilitates genetic manipulation inP. pastoris , providing high transformation efficiency and plasmid stability.IMPORTANCE Secretory production of recombinant proteins is the most important application of the methylotrophic yeastPichia pastoris , a species that permits mass production of heterologous proteins. To date, the genetic engineering ofP. pastoris has relied largely on integrative vectors due to the lack of user-friendly tools. Autonomously replicatingPichia plasmids are expected to facilitate genetic manipulation; however, the existing systems, which use autonomously replicating sequences (ARSs) such as theP. pastoris -specific ARS (PARS1 ), are known to be inherently unstable for plasmid replication and distribution. Recently, the centromeric DNA sequences ofP. pastoris were identified in back-to-back studies published by several groups; therefore, a new episomal plasmid vector with centromere DNA as a tool for genetic manipulation ofP. pastoris is ready to be developed.[Refereed]Scientific journal - May 2018, Microbial Cell Factories, 17, 76, EnglishMetabolome analysis-based design and engineering of a metabolic pathway in Corynebacterium glutamicum to match rates of simultaneous utilization of D-glucose and L-arabinose[Refereed]Scientific journal
- May 2018, Bioinformatics and Biology Insights, 12, 1 - 7, EnglishMathematical model for small size time series data of bacterial secondary metabolic pathways[Refereed]Scientific journal
- Apr. 2018, AMB Express, 8, 56, EnglishGenetic and physiological basis for antibody production by Kluyveromyces marxianus[Refereed]Scientific journal
- Mar. 2018, Journal of Bioscience and Bioengineering, 125(3) (3), 306 - 310, EnglishEnhanced cell-surface display of a heterologous protein using SED1 anchoring system in SED1-disrupted Saccharomyces cerevisiae strain[Refereed]Scientific journal
- Feb. 2018, Biotechnology for Biofuels, 11, 50, EnglishDirect and highly productive conversion of cyanobacteria Arthrospira platensis to ethanol with CaCl2 addition[Refereed]Scientific journal
- To efficiently utilize xylose, a major sugar component of hemicelluloses, in Saccharomyces cerevisiae requires the proper expression of varied exogenous and endogenous genes. To expand the repertoire of promoters in engineered xylose-utilizing yeast strains, we selected promoters in S. cerevisiae during cultivation and fermentation using xylose as a carbon source. To select candidate promoters that function in the presence of xylose, we performed comprehensive gene expression analyses using xylose-utilizing yeast strains both during xylose and glucose fermentation. Based on microarray data, we chose 29 genes that showed strong, moderate, and weak expression in xylose rather than glucose fermentation. The activities of these promoters in a xylose-utilizing yeast strain were measured by lacZ reporter gene assays over time during aerobic cultivation and microaerobic fermentation, both in xylose and glucose media. In xylose media, PTDH3, PFBA1, and PTDH1 were favorable for high expression, and PSED1, PHXT7, PPDC1, PTEF1, PTPI1, and PPGK1 were acceptable for medium–high expression in aerobic cultivation, and moderate expression in microaerobic fermentation. PTEF2 allowed moderate expression in aerobic culture and weak expression in microaerobic fermentation, although it showed medium–high expression in glucose media. PZWF1 and PSOL4 allowed moderate expression in aerobic cultivation, while showing weak but clear expression in microaerobic fermentation. PALD3 and PTKL2 showed moderate promoter activity in aerobic cultivation, but showed almost no activity in microaerobic fermentation. The knowledge of promoter activities in xylose cultivation obtained in this study will permit the control of gene expression in engineered xylose-utilizing yeast strains that are used for hemicellulose fermentation.Elsevier B.V., Jan. 2018, Journal of Bioscience and Bioengineering, 125(1) (1), 76 - 86, English[Refereed]Scientific journal
- Dec. 2017, Frontiers in Bioengineering and Biotechnology, 5, 81, EnglishImprovement of Xylose Fermentation Ability under Heat and Acid Co-Stress in Saccharomyces cerevisiae Using Genome Shuffling Technique[Refereed]Scientific journal
- The aim of this study was to improve biomass production of the green microalga Chlamydomonas sp. JSC4 under high salinity conditions. For this purpose, heavy ion beam-coupled mutagenesis and evolutionary engineering were performed using JSC4 as the parent strain. After long-term and continuous cultivation with high salinity, salt-resistant strains that grow well even in the presence of 7% sea salt were successfully obtained. Transcriptional analysis revealed inactivation of starch-to-lipid biosynthesis switching, which resulted in delayed starch degradation and decreased lipid content in the salt-resistant strains. Cellular aggregation and hypertrophy during high salinity were relieved in these strains, indicating strong resistance to salt stress. These results suggest that high salinity stress, not the salinity condition itself, is important for activating lipid accumulation mechanisms in microalgae. (C) 2017 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, Dec. 2017, BIORESOURCE TECHNOLOGY, 245(part B) (part B), 1484 - 1490, English[Refereed]Scientific journal
- Lipid from Chlamydomonas sp. JSC4 was used as a feedstock for biodiesel production. The lipid was found to contain high amounts of phospholipids and free fatty acid in addition to the triglycerides. Two enzymatic methods for the efficient conversion of the heterogenous lipid to fatty acid methyl esters (FAME) were carried out. The method using either a lipase cocktail containing Candida cylindracea lipase and Thermomyces lanuginosus lipase combination (m I) or immobilized Fusarium heterosporum lipase-expressing Aspergillus oryzae whole-cells (m II) were both successful. However, the method using lipase cocktail showed 30.8% relative stability after the fourth batch, whereas the whole-cell biocatalyst showed 98.1%. Although the whole-cell biocatalyst tolerated a wide range of water content, an exploration of the effect of water-methanol interaction on the biocatalytic process showed that 24% water and 7: 1 methanol to oil ratio is more favorable for FAME production. A higher initial methanol consumption rate facilitated a more stable system with the whole-cell biocatalyst, producing over 97% FAME in 32 h. The efficient conversion of a highly heterogenous substrate in the presence of high amounts of water could be an effective technique for the enzymatic conversion of microalgal lipids.ELSEVIER SCIENCE BV, Dec. 2017, ALGAL RESEARCH-BIOMASS BIOFUELS AND BIOPRODUCTS, 28, 16 - 23, English[Refereed]Scientific journal
- The yeast Saccharomyces cerevisiae cannot utilize xylose, but the introduction of a xylose isomerase that functions well in yeast will help overcome the limitations of the fungal oxido-reductive pathway. In this study, a diploid S. cerevisiae S288c[2n YMX12] strain was constructed expressing the Bacteroides thetaiotaomicron xylA (XI) and the Scheffersomyces stipitis xyl3 (XK) and the changes in the metabolite pools monitored over time. Cultivation on xylose generally resulted in gradual changes in metabolite pool size over time, whereas more dramatic fluctuations were observed with cultivation on glucose due to the diauxic growth pattern. The low G6P and F1,6P levels observed with cultivation on xylose resulted in the incomplete activation of the Crabtree effect, whereas the high PEP levels is indicative of carbon starvation. The high UDP-d-glucose levels with cultivation on xylose indicated that the carbon was channeled toward biomass production. The adenylate and guanylate energy charges were tightly regulated by the cultures, while the catabolic and anabolic reduction charges fluctuated between metabolic states. This study helped elucidate the metabolite distribution that takes place under Crabtree-positive and Crabtree-negative conditions when cultivating S. cerevisiae on glucose and xylose, respectively.SPRINGER HEIDELBERG, Oct. 2017, JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 44(10) (10), 1459 - 1470, English[Refereed]Scientific journal
- Aug. 2017, Scientific Reports, 7(1) (1), 8239, EnglishSynthesis of sulfo-sialic acid analogues: potent neuraminidase inhibitors in regards to anomeric functionality[Refereed]Scientific journal
- Aug. 2017, Scientific Reports, 7(1) (1), 8993, EnglishDevelopment of a comprehensive set of tools for genome engineering in a cold- and thermo-tolerant Kluyveromyces marxianus yeast strain[Refereed]Scientific journal
- Crystalline cellulose is one of the major contributors to the recalcitrance of lignocellulose to degradation, necessitating high dosages of cellulase to digest, thereby impeding the economic feasibility of cellulosic biofuels. Several recombinant cellulolytic yeast strains have been developed to reduce the cost of enzyme addition, but few of these strains are able to efficiently degrade crystalline cellulose due to their low cellulolytic activities. Here, by combining the cellulase ratio optimization with a novel screening strategy, we successfully improved the cellulolytic activity of a Saccharomyces cerevisiae strain displaying four different synergistic cellulases on the cell surface. The optimized strain exhibited an ethanol yield from Avicel of 57% of the theoretical maximum, and a 60% increase of ethanol titer from rice straw. To our knowledge, this work is the first optimization of the degradation of crystalline cellulose by tuning the cellulase ratio in a cellulase cell-surface display system. This work provides key insights in engineering the cellulase cocktail in a consolidated bioprocessing yeast strain. Biotechnol. Bioeng. 2017;114: 1201-1207. (c) 2017 Wiley Periodicals, Inc.WILEY, Jun. 2017, BIOTECHNOLOGY AND BIOENGINEERING, 114(6) (6), 1201 - 1207, English[Refereed]Scientific journal
- Apr. 2017, Scientific Reports, 4(7) (7), 45471, EnglishDynamic metabolic profiling together with transcription analysis reveals salinity-induced starch-to-lipid biosynthesis in alga Chlamydomonas sp. JSC4[Refereed]Scientific journal
- Mar. 2017, Biotechnology Journal, 12(3) (3), 1600400, EnglishSimultaneous conversion of free fatty acids and triglycerides to biodiesel by immobilized Aspergillus oryzae expressing Fusarium heterosporum lipase[Refereed]Scientific journal
- The surface of yeast cells has been an attractive interface for the effective use of cellulose. Surface enzymes, however, are difficult to visualize and evaluate. In this study, two kinds of unique anchoring regions were used to display the cellulase, endoglucanase (EG), on a yeast cell surface. Differences in the display level and the localization of EG were observed by atomic force microscopy. By surveying the yeast cell surface with a chemically modified cantilever, the interactive force between the cellulose and EG was measured. Force curve mapping revealed differences in the display levels and the localization of EG according to anchoring regions. The proposed methodology enables visualization of displayed enzymes such as EG on the yeast cell surface. (C) 2016 Elsevier B.V. All rights reserved.ELSEVIER SCIENCE BV, Mar. 2017, COLLOIDS AND SURFACES B-BIOINTERFACES, 151, 134 - 142, English[Refereed]Scientific journal
- Mar. 2017, Microbial Cell Factories, 16, 44, EnglishEnzymatic improvement of mitochondrial thiol oxidase Erv1 for oxidized glutathione fermentation by Saccharomyces cerevisiae[Refereed]Scientific journal
- Mar. 2017, Scientific Reports, 7, 43518, EnglishA systematic approach to time-series metabolite profiling and RNA-seq analysis of Chinese hamster ovary cell culture[Refereed]Scientific journal
- This work illustrates several theoretical fundamentals for the application of THz vibrational spectroscopy to molecular characterization in the solid state using two different types of saccharide systems as examples. Four subjects have been specifically addressed: (1) the qualitative differences in the molecular vibrational signatures monitored by THz and mid-IR vibrational spectroscopy; (2) the selection rules for THz vibrational spectroscopy as applied to crystalline and amorphous systems; (3) a normal mode simulation, using a-l-xylose as an example; and (4) a rigorous mode analysis to quantify the percentage contributions of the intermolecular and intramolecular vibrations to the normal mode of interest.WILEY-V C H VERLAG GMBH, Feb. 2017, CHEMISTRY-AN ASIAN JOURNAL, 12(3) (3), 324 - 331, English[Refereed]Scientific journal
- Feb. 2017, Shimadzu Journal, 5(1) (1), 17 - 21, EnglishApplication of LC-MS/MS analysis for time-lapse amino acid metabolomics in CHO cell culture.[Refereed]Scientific journal
- Volume 3, no. 2, e00389-15, 2015. Page 1, column 1, lines 26 and 27: "silencing mediator for retinoic acid and thyroid hormone receptor" should read "singlemolecule real-time."American Society for Microbiology, 2017, Genome Announcements, 5(5) (5), English[Refereed]Scientific journal
- Jan. 2017, Nature Communications, 8, 14007, EnglishA RuBisCO-mediated carbon metabolic pathway in methanogenic archaea[Refereed]Scientific journal
- Dec. 2016, Frontiers in Microbiology, 7, 2050, EnglishSuccinate and lactate production from Euglena gracilis during dark, anaerobic conditions.[Refereed]Scientific journal
- The feedstocks used for the production of bio-based chemicals have recently expanded from edible sugars to inedible and more recalcitrant forms of lignocellulosic biomass. To produce biobased chemicals from renewable polysaccharides, several bioprocessing approaches have been developed and include separate hydrolysis and fermentation (SHF), simultaneous saccharification and fermentation (SSF), and consolidated bioprocessing (CBP). In the last decade, SHF, SSF, and CBP have been used to generate macromolecules and aliphatic and aromatic compounds that are capable of serving as sustainable, drop-in substitutes for petroleum-based chemicals. The present review focuses on recent progress in the bioprocessing of microbially produced chemicals from renewable feedstocks, including starch and lignocellulosic biomass. In particular, the technological feasibility of bio-based chemical production is discussed in terms of the feedstocks and different bioprocessing approaches, including the consolidation of enzyme production, enzymatic hydrolysis of biomass, and fermentation.CURRENT BIOLOGY LTD, Dec. 2016, CURRENT OPINION IN BIOTECHNOLOGY, 42, 30 - 39, English[Refereed]Scientific journal
- Wiley-VCH Verlag, Nov. 2016, Industrial Biotechnology: Products and Processes, 161 - 185, English[Refereed]In book
- Xylem vessels, the water-conducting cells in vascular plants, undergo characteristic secondary wall deposition and programmed cell death. These processes are regulated by the VASCULAR-RELATED NAC-DOMAIN (VND) transcription factors. Here, to identify changes in metabolism that occur during protoxylem vessel element differentiation, we subjected tobacco (Nicotiana tabacum) BY-2 suspension culture cells carrying an inducible VND7 system to liquid chromatography-mass spectrometry-based wide-target metabolome analysis and transcriptome analysis. Time-course data for 128 metabolites showed dynamic changes in metabolites related to amino acid biosynthesis. The concentration of glyceraldehyde 3-phosphate, an important intermediate of the glycolysis pathway, immediately decreased in the initial stages of cell differentiation. As cell differentiation progressed, specific amino acids accumulated, including the shikimate-related amino acids and the translocatable nitrogen-rich amino acid arginine. Transcriptome data indicated that cell differentiation involved the active up-regulation of genes encoding the enzymes catalyzing fructose 6-phosphate biosynthesis from glyceraldehyde 3-phosphate, phosphoenolpyruvate biosynthesis from oxaloacetate, and phenylalanine biosynthesis, which includes shikimate pathway enzymes. Concomitantly, active changes in the amount of fructose 6-phosphate and phosphoenolpyruvate were detected during cell differentiation. Taken together, our results show that protoxylem vessel element differentiation is associated with changes in primary metabolism, which could facilitate the production of polysaccharides and lignin monomers and, thus, promote the formation of the secondary cell wall. Also, these metabolic shifts correlate with the active transcriptional regulation of specific enzyme genes. Therefore, our observations indicate that primary metabolism is actively regulated during protoxylem vessel element differentiation to alter the cell's metabolic activity for the biosynthesis of secondary wall polymers.AMER SOC PLANT BIOLOGISTS, Nov. 2016, PLANT PHYSIOLOGY, 172(3) (3), 1612 - 1624, English[Refereed]Scientific journal
- Examination of the effects of high temperature on the photoinhibition of photosystem II (PSII) in the cyanobacterium Synechocystis sp. PCC 6803 revealed that the extent of photoinhibition of PSII was lower at moderately high temperatures (35-42 degrees C) than at 30 degrees C. Photodamage to PSII, as determined in the presence of chloramphenicol, which blocks the repair of PSII, was accelerated at the moderately high temperatures but the effects of repair were greater than those of photodamage. The synthesis de novo of the D1 protein, which is essential for the repair of PSII, was enhanced at 38 degrees C. Electron transport and the synthesis of ATP were also enhanced at 38 degrees C, while levels of reactive oxygen species fell. Inhibition of the Calvin-Benson cycle with glycolaldehyde abolished the enhancement of repair of PSII at 38 degrees C, suggesting that an increase in the activity of the Calvin-Benson cycle might be required for the enhancement of repair at moderately high temperatures. The synthesis de novo of metabolic intermediates of the Calvin-Benson cycle, such as 3-phosphoglycerate, was also enhanced at 38 degrees C. We propose that moderate heat stress might enhance the repair of PSII by stimulating the synthesis of ATP and depressing the production of reactive oxygen species, via the stimulation of electron transport and suppression of the accumulation of excess electrons on the acceptor side of photosystem I, which might be driven by an increase in the activity of the Calvin-Benson cycle.OXFORD UNIV PRESS, Nov. 2016, PLANT AND CELL PHYSIOLOGY, 57(11) (11), 2417 - 2426, English[Refereed]Scientific journal
- Nov. 2016, Applied Spectroscopy, 71(3) (3), 463 - 471, EnglishHigh-speed scanning for the quantitative evaluation of glycogen concentration in bioethanol feedstock Synechocystis sp. PCC6803 by a near-infrared hyperspectral imaging system with a new near-infrared spectral camera.[Refereed]Scientific journal
- Recombinant yeast strains displaying aheterologous cellulolytic enzymes on their cell surfaces using a glycosylphosphatidylinositol (GPI) anchoring system are a promising strategy for bioethanol production from lignocellulosic materials. A crucial step for cell wall localization of the enzymes is the intracellular transport of proteins in yeast cells. Therefore, the addition of a highly efficient secretion signal sequence is important to increase the amount of the enzymes on the yeast cell surface. In this study, we demonstrated the effectiveness of a novel signal peptide (SP) sequence derived from the Saccharomyces cerevisiae SED1 gene for cell-surface display and secretory production of cellulolytic enzymes. Gene cassettes with SP sequences derived fromS. cerevisiae SED1 (SED1SP), Rhizopus oryzae glucoamylase (GLUASP), and S. cerevisiae alpha-mating pheromone (MF alpha 1SP) were constructed for cell-surface display of Aspergillus aculeatus beta-glucosidase (BGL1) and Trichoderma reesei endoglucanase II (EGII). These gene cassettes were integrated into the S. cerevisiae genome. The recombinant strains with the SED1SP showed higher cell-surface BGL and EG activities than those with the conventional SP sequences (GLUASP and MF alpha 1SP). The novel SP sequence also improved the secretory production of BGL and EG in S. cerevisiae. The extracellular BGL activity of the recombinant strains with the SED1SP was 1.3- and 1.9-fold higher than the GLUASP and MF alpha 1SP strains, respectively. Moreover, the utilization of SED1SP successfully enhanced the secretory production of BGL in Pichia pastoris. The utilization of the novel SP sequence is a promising option for highly efficient cell-surface display and secretory production of heterologous proteins in various yeast species. (C) 2016 Wiley Periodicals, Inc.WILEY-BLACKWELL, Nov. 2016, BIOTECHNOLOGY AND BIOENGINEERING, 113(11) (11), 2358 - 2366, English[Refereed]Scientific journal
- N-acetyl-D-glucosamine (GlcNAc) is the building block of chitin, which is one of the most abundant renewable resources in nature after cellulose. Therefore, a microorganism that can utilize GlcNAc is necessary for chitin-based biorefinery. In this study, we report on the screening and characterization of yeast strains for bioethanol production from GlcNAc. We demonstrate that Scheffersomyces (Pichia) stipitis strains can use GlcNAc as the sole carbon source and produce ethanol. S. stipitis NBRC1687, 10007, and 10063 strains consumed most of the 50 g/L GlcNAc provided, and produced 14.5 +/- 0.6, 15.0 +/- 0.3, and 16.4 +/- 0.3 g/L of ethanol after anaerobic fermentation at 30 degrees C for 96 h. The ethanol yields of these strains were approximately 81, 75, and 82 % (mol ethanol/mol GlcNAc consumed), respectively. Moreover, S. stipitis NBRC10063 maintained high GlcNAc-utilizing capacity at 35 degrees C, and produced 12.6 +/- 0.7 g/L of ethanol after 96 h. This strain also achieved the highest ethanol titer (23.3 +/- 1.0 g/L) from 100 g/L GlcNAc. To our knowledge, this is the first report on ethanol production via fermentation of GlcNAc by naturally occurring yeast strains.BIOMED CENTRAL LTD, Oct. 2016, AMB EXPRESS, 6, 83, English[Refereed]Scientific journal
- Waste biorefinery aims to maximize the output of value-added products from various artificial/agricultural wastes by using integrated bioprocesses. To make waste biorefinery economically feasible, it is thus necessary to develop a low-cost, environment-friendly technique to perform simultaneous biodegradation and bioconversion of waste materials. Cell-surface display engineering is a novel, cost-effective technique that can auto-immobilize proteins on the cell exterior of microorganisms, and has been applied for use with waste biofinery. Through tethering different enzymes (e.g., cellulase, lipase, and protease) or metal-binding peptides on cell surfaces, various yeast strains can effectively produce biofuels and biochemicals from sugar/protein-rich waste materials, catalyze waste oils into biodiesels, or retrieve heavy metals from wastewater. This review critically summarizes recent applications of yeast cell-surface display on various types of waste biorefineries, highlighting its potential and future challenges with regard to commercializing this technology. (C) 2016 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, Sep. 2016, BIORESOURCE TECHNOLOGY, 215, 324 - 333, English[Refereed]Scientific journal
- Cyanobacterial hydrogenases are important owing to the association between hydrogen metabolism and cell physiology and the production of future renewable energy. Many studies have examined hydrogen productivity, transcriptional regulation of hydrogenases, and the biochemistry of hydrogenases; however the relationship between hydrogen and primary carbon metabolism using metabolomic techniques has not been elucidated. Here, we studied the effect of the genetic manipulation of a hydrogenase on primary carbon metabolism in the model unicellular cyanobacterium Synechocystis sp. PCC 6803. Metabolomic analysis revealed that the hoxH mutant with reduced hoxH transcripts exhibited increased sugar phosphates in dark, anaerobic conditions. Organic acids, lactate, succinate, fumarate, and malate increased substantially by the hoxH mutation both inside and outside of cells in dark, anaerobic conditions. Transcriptome analysis revealed higher expression of genes encoding the RNA polymerase sigma factor SigE, which is a positive regulator of sugar catabolism, and 6-phosphogluconate dehydrogenase in the hoxH mutant than in the wild-type strain. Immunoblotting results showed that sugar catabolic enzymes and SigE proteins increased in the hoxH mutant. These results demonstrate the wide alterations of primary metabolism by the genetic manipulation of a hydrogenase subunit in this cyanobacterium. (C) 2016 Elsevier B.V. All rights reserved.ELSEVIER SCIENCE BV, Sep. 2016, ALGAL RESEARCH-BIOMASS BIOFUELS AND BIOPRODUCTS, 18, 305 - 313, English[Refereed]Scientific journal
- Biofuels from microalgae is now a hot issue of great potential. However, achieving high starch productivity with photoautotrophic microalgae is still challenging. A feasible approach to enhance the growth and target product of microalgae is to conduct mixotrophic cultivation. The appropriate acetate addition combined with CO2 supply as dual carbon sources (i.e., mixotrophic cultivation) could enhance the cell growth of some microalgae species, but the effect of acetate-mediated mixotrophic culture mode on carbohydrate accumulation in microalgae remains unclear. Moreover, there is still lack of the information concerning how to increase the productivity of carbohydrates from microalgae under acetate-amended mixotrophic cultivation and how to optimize the engineering strategies to achieve the goal. This study was undertaken to develop an optimal acetate-contained mixotrophic cultivation system coupled with effective operation strategies to markedly improve the carbohydrate productivity of Chlorella sorokiniana NIES-2168. The optimal carbohydrate productivity of 695 mg/L/d was obtained, which is the highest value ever reported. The monosaccharide in the accumulated carbohydrates is mainly glucose (i.e., 85-90%), which is very suitable for bio-alcohols fermentation. Hence, by applying the optimal process developed in this study, C. sorokiniana NIES-2168 has a high potential to serve as a feedstock for subsequent biofuels conversion.WILEY-V C H VERLAG GMBH, Aug. 2016, BIOTECHNOLOGY JOURNAL, 11(8) (8), 1072 - 1081, English[Refereed]Scientific journal
- The presence of phospholipid has been a challenge in liquid enzymatic biodiesel production. Among six lipases that were screened, lipase AY had the highest hydrolysis activity and a competitive transesterification activity. However, it yielded only 21.1% FAME from oil containing phospholipids. By replacing portions of these lipases with a more robust bioFAME lipase, CalT, the combination of lipase AY-CalT gave the highest FAME yield with the least amounts of free fatty acids and partial glycerides. A higher methanol addition rate reduced FAME yields for lipase DF-CalT and A10D-CalT combinations while that of lipase AY-CalT combination improved. Optimizing the methanol addition rate for lipase AY-CalT resulted in a FAME yield of 88.1% at 2 h and more than 95% at 6 h. This effective use of lipases could be applied for the rapid and economic conversion of unrefined oils to biodiesel. (C) 2016 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, Jul. 2016, BIORESOURCE TECHNOLOGY, 211, 224 - 230, English[Refereed]Scientific journal
- May 2016, Metabolic Engineering Communications, 3, 130 - 141, EnglishImproved sugar-free succinate production by Synechocystis sp. PCC 6803 following identification of the limiting steps in glycogen catabolism.[Refereed]Scientific journal
- Apr. 2016, Scientific Reports, 6, 24550, EnglishEngineering of a novel cellulose-adherent cellulolytic Saccharomyces cerevisiae for cellulosic biofuel production.[Refereed]Scientific journal
- Xylitol, a value-added polyol deriving from D-xylose, is widely used in both the food and pharmaceutical industries. Despite extensive studies aiming to streamline the production of xylitol, the manufacturing cost of this product remains high while demand is constantly growing worldwide. Biotechnological production of xylitol from lignocellulosic waste may constitute an advantageous and sustainable option to address this issue. However, to date, there have been few reports of biomass conversion to xylitol. In the present study, xylitol was directly produced from rice straw hydrolysate using a recombinant Saccharomyces cerevisiae YPH499 strain expressing cytosolic xylose reductase (XR), along with beta-glucosidase (BGL), xylosidase (XYL), and xylanase (XYN) enzymes (co-) displayed on the cell surface; xylitol production by this strain did not require addition of any commercial enzymes. All of these enzymes contributed to the consolidated bioprocessing (CBP) of the lignocellulosic hydrolysate to xylitol to produce 5.8 g/L xylitol with 79.5 % of theoretical yield from xylose contained in the biomass. Furthermore, nanofiltration of the rice straw hydrolysate provided removal of fermentation inhibitors while simultaneously increasing sugar concentrations, facilitating high concentration xylitol production (37.9 g/L) in the CBP. This study is the first report (to our knowledge) of the combination of cell surface engineering approach and membrane separation technology for xylitol production, which could be extended to further industrial applications.SPRINGER, Apr. 2016, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 100(8) (8), 3477 - 3487, English[Refereed]Scientific journal
- Lignocellulosic hydrolysates contain compounds that inhibit microbial growth and fermentation, thereby decreasing the productivity of biofuel and biochemical production. In particular, the heterocyclic aldehyde furfural is one of the most toxic compounds found in these hydrolysates. We previously demonstrated that Corynebacterium glutamicum converts furfural into the less toxic compounds furfuryl alcohol and 2-furoic acid. To date, however, the genes involved in these oxidation and reduction reactions have not been identified in the C. glutamicum genome. Here, we show that Cgl0331 (designated FudC) is mainly responsible for the reduction of furfural into furfuryl alcohol in C. glutamicum. Deletion of the gene encoding FudC markedly diminished the in vivo reduction of furfural to furfuryl alcohol. Purified His-tagged FudC protein from Escherichia coli was also shown to convert furfural into furfuryl alcohol in an in vitro reaction utilizing NADPH, but not NADH, as a cofactor. Kinetic measurements demonstrated that FudC has a high affinity for furfural but has a narrow substrate range for other aldehydes compared to the protein responsible for furfural reduction in E. coli.SPRINGER, Mar. 2016, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 100(6) (6), 2685 - 2692, English[Refereed]Scientific journal
- Mar. 2016, AMB Express, 6(1) (1), 4, EnglishDisruption of PHO13 improves ethanol production via the xylose isomerase pathway[Refereed]Scientific journal
- Sorghum bagasse pretreated with diluted acid, which was predominantly composed of glucan (59%) and xylan (7.2%), was used as a lignocellulosic feedstock to produce D-phenyllactic acid (PhLA) by a recombinant Escherichia coli strain expressing phenylpyruvate reductase from Wickerhamia fluorescens. Compared to filter paper hydrolysate, the PhLA yield was reduced by 35% during fermentation with enzymatic hydrolysate of sorghum bagasse as a carbon source, and metabolomics analysis revealed that intracellular levels of erythrose-4-phosphate and phosphoenolpyruvate and NAD(P)H regeneration for PhLA production from glucose markedly reduced. Compared to the separate hydrolysis and fermentation (SHF) with sorghum bagasse hydrolysate, simultaneous saccharification and fermentation (SSF) of sorghum bagasse under glucose limitation conditions yielded 4.8-fold more PhLA with less accumulation of eluted components of p-coumaric acid and aldehydes, which inhibited PhLA fermentation. These results suggest that gradual hydrolysis of sorghum bagasse during SSF reduces the accumulation of both glucose and fermentation inhibitors, collectively leading to increased PhLA yield.The Japan Institute of Energy, Jan. 2016, Proceedings of the Conference on Biomass Science, 11, 35 - 36, Japanese[Refereed]International conference proceedings
- Improving the production of ethanol from xylose is an important goal in metabolic engineering of Saccharomyces cerevisiae. Furthermore, S. cerevisiae must produce ethanol in the presence of weak acids (formate and acetate) generated during pre-treatment of lignocellulosic biomass. In this study, weak acid-containing xylose fermentation was significantly improved using cells that were acclimated to the weak acids during pre-cultivation. Transcriptome analyses showed that levels of transcripts for transcriptional/translational machinery-related genes (RTC3 and ANB1) were enhanced by formate and acetate acclimation. Recombinant yeast strains overexpressing RTC3 and ANB1 demonstrated improved ethanol production from xylose in the presence of the weak acids, along with improved tolerance to the acids. Novel metabolic engineering strategy based on the combination of short-term acclimation and system-wide analysis was developed, which can develop stress-tolerant strains in a short period of time, although conventional evolutionary engineering approach has required long periods of time to isolate inhibitor-adapted strains.SPRINGER, Jan. 2016, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 100(2) (2), 1027 - 1038, English[Refereed]Scientific journal
- The presence of phospholipids in oil has been a major hurdle in the production of biodiesel using immobilized Aspergillus oryzae whole-cell biocatalysts. A density of phospholipids within the range of 10-30% could reduce both the rate of production and the final yield of biodiesel. Phospholipids in the oil leads to the formation of water-in-oil phospholipid-based reverse micelles. The water that activates the enzymatic process is observed to be trapped inside these reverse micelles. This has resulted in the inactivation of the reaction systems and has subsequently led to the deactivation of the immobilized lipase by the extended residence time of the added methanol. A reaction system involving gentle agitation and higher amount of water was found to reduce the reverse micelles formation. This simple technique improved the conversion efficiency by approximately 3-folds, producing a final biodiesel of more than 90%, using immobilized A. oryzae whole cells expressing Fusarium heterosporum lipase. This demonstrates that, the above technique could be successfully applied to the enzymatic biodiesel conversion of oils containing high amounts of phospholipids such as that from microalgae. (C) 2015 Elsevier B.V. All rights reserved.ELSEVIER SCIENCE BV, Jan. 2016, BIOCHEMICAL ENGINEERING JOURNAL, 105, 10 - 15, English[Refereed]Scientific journal
- Conferring biomass hydrolysis activity on yeast through genetic engineering has paved the way for the development of groundbreaking processes for producing liquid fuels and commodity chemicals from lignocellulosic biomass. However, the overproduction and misfolding of heterologous and endogenous proteins can trigger cellular stress, increasing the metabolic burden and retarding growth. Improving the efficiency of lignocellulosic breakdown requires engineering of yeast secretory pathway based on system-wide metabolic analysis as well as DNA constructs for enhanced cellulase gene expression with advanced molecular biology tools. Also, yeast is subjected to severe stress due to toxic compounds generated during lignocellulose pretreatment in consolidated saccharification and fermentation processes. The prospect for development of robust yeast strains makes combining evolutionary and rational engineering strategies.ELSEVIER SCI LTD, Dec. 2015, CURRENT OPINION IN CHEMICAL BIOLOGY, 29, 1 - 9, English[Refereed]Scientific journal
- Genetic engineering and metabolite profiling for the overproduction of polyhydroxybutyrate (PHB), which is a carbon material in biodegradable plastics, were examined in the unicellular cyanobacterium Synechocystis sp. PCC 6803. Transconjugants harboring cyanobacterial expression vectors that carried the pha genes for PHB biosynthesis were constructed. The overproduction of PHB by the engineering cells was confirmed through microscopic observations using Nile red, transmission electron microscopy (TEM), or nuclear magnetic resonance (NMR). We successfully recovered PHB from transconjugants prepared from nitrogen-depleted medium without sugar supplementation in which PHB reached approximately 7% (w/w) of the dry cell weight, showing a value of 12-fold higher productivity in the transconjugant than that in the control strain. We also measured the intracellular levels of acetyl-CoA, acetoacetyl-CoA, and 3-hydroxybutyryl-CoA (3HB-CoA), which are intermediate products for PHB. The results obtained indicated that these products were absent or at markedly low levels when cells were subjected to the steady-state growth phase of cultivation under nitrogen depletion for the overproduction of bioplastics. Based on these results, efficient factors were discussed for the overproduction of PHB in recombinant cyanobacteria. (C) 2015, The Society for Biotechnology, Japan. All rights reserved.SOC BIOSCIENCE BIOENGINEERING JAPAN, Nov. 2015, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 120(5) (5), 510 - 517, English[Refereed]Scientific journal
- Nitrogen starvation is known to induce the accumulation of triacylglycerol (TAG) in many microalgae, and potential use of microalgae as a source of biofuel has been explored. However, nitrogen starvation also stops cellular growth. The expression of cyanobacterial acyl-acyl carrier protein (ACP) reductase in the unicellular red alga Cyanidioschyzon merolae chloroplasts resulted in an accumulation of TAG, which led to an increase in the number and size of lipid droplets while maintaining cellular growth. Transcriptome and metabolome analyses showed that the expression of acyl-ACP reductase altered the activities of several metabolic pathways. The activities of enzymes involved in fatty acid synthesis in chloroplasts, such as acetyl-CoA carboxylase and pyruvate dehydrogenase, were up-regulated, while pyruvate decarboxylation in mitochondria and the subsequent consumption of acetyl-CoA by the tricarboxylic acid (TCA) cycle were down-regulated. Aldehyde dehydrogenase, which oxidizes fatty aldehydes to fatty acids, was also up-regulated in the acyl-ACP reductase expresser. This activation was required for the lipid droplet accumulation and metabolic changes observed in the acyl-ACP reductase expresser. Nitrogen starvation also resulted in lipid droplet accumulation in C. merolae, while cell growth ceased as in the case of other algal species. The metabolic changes that occur upon the expression of acyl-ACP reductase are quite different from those caused by nitrogen starvation. Therefore, there should be a method for further increasing the storage lipid level while still maintaining cell growth that is different from the metabolic response to nitrogen starvation.OXFORD UNIV PRESS, Oct. 2015, PLANT AND CELL PHYSIOLOGY, 56(10) (10), 1962 - 1980, English[Refereed]Scientific journal
- Oct. 2015, 化学と生物, 53(10) (10), 689 - 695, Japaneseバイオリファイナリーの現状と展望Scientific journal
- Sep. 2015, 生物工学会誌, 93(9) (9), 523 - 526, Japanese合成生物工学によるモノづくり微生物のデザインに向けてScientific journal
- Sep. 2015, Biotechnology for Biofuels, 8, 162, EnglishCombined cell-surface display- and secretion-based strategies for production of cellulosic ethanol with Saccharomyces cerevisiae[Refereed]Scientific journal
- Photosynthetic microorganisms, such as cyanobacteria and microalgae, are currently being investigated as alternative biomass resources for bioethanol production, owing to their benefits, including high-photosynthetic activity and whole-year cultivation without utilization of arable land. Polyglucans comprise the major carbohydrate content of these organisms. Polyglucans can be utilized as a carbon source for microbial fermentation. Although polyglucan production has so far been promoted by nutrient limitation, it must be further enhanced to accommodate market demand. This review focuses on the recent progress in the production of -polyglucans such asglycogen and starch in cyanobacteria and green microalgae via cultivation design, including modifying the nutrient supply and replacing the growth medium. The control and manipulation of polyglucan metabolism necessitates the elucidation of the polyglucan production mechanism. We reviewed gene expression and metabolite accumulation profiles of cyanobacteria and green microalgae during nutrient limitation-stimulated -polyglucan accumulation. We also focus on the enhancement in cyanobacterial glycogen production via the genetic engineering of glycolysis, CO2 concentration mechanism, and photosynthetic light-harvesting protein based on the polyglucan accumulation mechanism. The combined strategies of cultivation design and genetic engineering should be considered for further enhancement of polyglucan productivity for bioethanol production.WILEY-V C H VERLAG GMBH, Jun. 2015, BIOTECHNOLOGY JOURNAL, 10(6) (6), 886 - 898, English[Refereed]Scientific journal
- Jun. 2015, Biotechnology for Biofuels, 8, 88, EnglishPrecipitate obtained following membrane separation of hydrothermally pretreated rice straw liquid revealed by 2D NMR to have high lignin content[Refereed]Scientific journal
- Mechanical milling and membrane separation were applied to simultaneous saccharification and co-fermentation from hydrothermally pretreated rice straw. Mechanical milling with minimized 4 cycles enabled 37.5 +/- 3.4 g L-1 and 45.3 +/- 4.4 g L-1 of ethanol production after 48 h by xylose-fermenting Saccharomyces cerevisiae from solid fractions (200 and 250 g L-1) of pretreated rice straw with 5 filter paper unit g-biomass(-1) cellulase (respectively, 77.3 +/- 7.1% and 74.7 +/- 7.3% of theoretical ethanol yield). Use of a membrane-based process including nanofiltration and ultrafiltration increased the sugar concentrations in the liquid fraction of pretreated rice straw and addition of this liquid fraction to 250 g L-1 solid fraction increased ethanol production to 52.0 +/- 0.4 g L-1 (73.8 +/- 0.6% of theoretical ethanol yield). Mechanical milling was effective in increasing enzymatic hydrolysis of the solid fraction and membrane separation steps increased the ethanol titer during co-fermentation, leading to a proposal for combining these processes for ethanol production from whole rice straw. (C) 2015 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, Jun. 2015, BIORESOURCE TECHNOLOGY, 185, 263 - 268, English[Refereed]Scientific journal
- May 2015, バイオインダストリー, 32(5) (5), 26 - 31, JapaneseCBPによるバイオエタノール生産技術の開発Scientific journal
- Dilute acid-pretreated sorghum bagasse, which was predominantly composed of glucan (59%) and xylose (7.2%), was used as a lignocellulosic feedstock for D-phenyllactic acid (PhLA) production by a recombinant Escherichia coli strain expressing phenylpyruvate reductase from Wickerhamia fluorescens. During fermentation with enzymatic hydrolysate of sorghum bagasse as a carbon source, the PhLA yield was reduced by 35% compared to filter paper hydrolysate, and metabolomics analysis revealed that NAD(P)H regeneration and intracellular levels of erythrose-4-phosphate and phosphoenolpyruvate for PhLA biosynthesis markedly reduced. Compared to separate hydrolysis and fermentation (SHF) with sorghum bagasse hydrolysate, simultaneous saccharification and fermentation (SSF) of sorghum bagasse under glucose limitation conditions yielded 4.8-fold more PhLA with less accumulation of eluted components, including p-coumaric acid and aldehydes, which inhibited PhLA fermentation. These results suggest that gradual enzymatic hydrolysis during SSF enhances PhLA production under glucose limitation and reduces the accumulation of fermentation inhibitors, collectively leading to increased PhLA yield. (c) 2015 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, Apr. 2015, BIORESOURCE TECHNOLOGY, 182, 169 - 178, English[Refereed]Scientific journal
- Apr. 2015, Microbial Cell Factories, 14, 56, EnglishEvaluation of genes involved in oxidative phosphorylation in yeast by developing a simple and rapid method to measure mitochondrial ATP synthetic activity.[Refereed]Scientific journal
- Apr. 2015, Genome Announcements, 3(2) (2), e00389 - 15, EnglishComplete genome sequence of Kluyveromyces marxianus NBRC1777, a nonconventional thermotolerant yeast[Refereed]Scientific journal
- Mar. 2015, 生物工学会誌, 93(3) (3), 122 - 129, Japanese代謝プロファイリングに基づく微生物育種技術の開発と応用Scientific journal
- Recent increasing attention to environmental issues and the shortage of oil resources have spurred political and industrial interest in the development of environmental friendly and cost-effective processes for the production of bio-based chemicals from renewable resources. Thus, microbial production of commercially important chemicals is viewed as a desirable way to replace current petrochemical production. Corynebacterium glutamicum, a Gram-positive soil bacterium, is one of the most important industrial microorganisms as a platform for the production of various amino acids. Recent research has explored the use of C. glutamicum as a potential cell factory for producing organic acids such as lactate and succinate, both of which are commercially important bulk chemicals. Here, we summarize current understanding in this field and recent metabolic engineering efforts to develop C. glutamicum strains that efficiently produce L- and D-lactate, and succinate from renewable resources.SPRINGER HEIDELBERG, Mar. 2015, JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 42(3) (3), 375 - 389, English[Refereed]Scientific journal
- The hydrolysis of lignocellulosic biomass liberates sugars, primarily glucose and xylose, which are subsequently converted to ethanol by microbial fermentation. The rapid and efficient fermentation of xylose by recombinant Saccharomyces cerevisiae strains is limited by weak acids generated during biomass pretreatment processes. In particular, acetic acid negatively affects cell growth, xylose fermentation rate, and ethanol production. The ability of S. cerevisiae to efficiently utilize xylose in the presence of acetic acid is an essential requirement for the cost-effective production of ethanol from lignocellulosic hydrolysates. Here, an acetic acid-responsive transcriptional activator, HAA1, was overexpressed in a recombinant xylose-fermenting S. cerevisiae strain to yield BY4741X/HAA1. This strain exhibited improved cell growth and ethanol production from xylose under aerobic and oxygen limited conditions, respectively, in the presence of acetic acid. The HAA1p regulon enhanced transcript levels in BY4741X/HAA1. The disruption of PHO13, a p-nitrophenylphosphatase gene, in BY4741X/HAA1 led to further improvement in both yeast growth and the ability to ferment xylose, indicating that HAM overexpression and PHO13 deletion act by different mechanisms to enhance ethanol production. (C) 2014, The Society for Biotechnology, Japan. All rights reserved.SOC BIOSCIENCE BIOENGINEERING JAPAN, Mar. 2015, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 119(3) (3), 297 - 302, English[Refereed]Scientific journal
- Mar. 2015, Biotechnology for Biofuels, 8, 48, EnglishDynamic metabolic profiling of the marine microalga Chlamydomonas sp. JSC4 and enhancing its oil production by optimizing light intensity[Refereed]Scientific journal
- Feb. 2015, BMC Microbiology, 15, 43, EnglishHyperphosphorylation of DegU cancels CcpA-dependent catabolite repression of rocG in Bacillus subtilis[Refereed]Scientific journal
- Recombinant yeast strains that display heterologous amylolytic enzymes on their cell surface via the glycosylphosphatidylinositol (GPI)-anchoring system are considered as promising biocatalysts for direct ethanol production from starchy materials. For the effective hydrolysis of these materials, the ratio optimization of multienzyme activity displayed on the cell surface is important. In this study, we have presented a ratio control system of multienzymes displayed on the yeast cell surface by using different GPI-anchoring domains. The novel gene cassettes for the cell-surface display of Streptococcus bovis alpha-amylase and Rhizopus oryzae glucoamylase were constructed using the Saccharomyces cerevisiae SED1 promoter and two different GPI-anchoring regions derived from Saccharomyces cerevisiae SED1 or SAG1. These gene cassettes were integrated into the Saccharomyces cerevisiae genome in different combinations. Then, the cell-surface alpha-amylase and glucoamylase activities and ethanol productivity of these recombinant strains were evaluated. The combinations of the gene cassettes of these enzymes affected the ratio of cell-surface alpha-amylase and glucoamylase activities and ethanol productivity of the recombinant strains. The highest ethanol productivity from raw starch was achieved by the strain harboring one alpha-amylase gene cassette carrying the SED1-anchoring region and two glucoamylase gene cassettes carrying the SED1-anchoring region (BY-AASS/GASS/GASS). This strain yielded 22.5 +/- 0.6 g/L of ethanol from 100 g/L of raw starch in 120 h of fermentation.SPRINGER, Feb. 2015, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 99(4) (4), 1655 - 1663, English[Refereed]Scientific journal
- The mechanisms of how zinc protects the cells against acetic acid toxicity and acts as an antioxidant are still not clear. Here we present results of the metabolic profiling of the eukaryotic model yeast species Saccharomyces cerevisiae subjected to long term high concentration acetic acid stress treatment in the presence and absence of zinc supplementation. Zinc addition decreased the release of reactive oxygen species (ROS) in the presence of chronic acetic acid stress. The dynamic changes in the accumulation of intermediates in central carbon metabolism were observed, and higher contents of intracellular alanine, valine and serine were observed by zinc supplementation. The most significant change was observed in alanine content, which is 3.51-fold of that of the control culture in cells in the stationary phase. Subsequently, it was found that 0.5 g L-1 alanine addition resulted in faster glucose consumption in the presence of 5 g L-1 acetic acid, and apparently decreased ROS accumulation in zinc-supplemented cells. This indicates that alanine exerted its antioxidant effect at least partially through the detoxification of acetic acid. In addition, intracellular glutathione (GSH) accumulation was enhanced by zinc addition, which is related to the protection of yeast cells from the oxidative injury caused by acetic acid. Our studies revealed for the first time that zinc modulates cellular amino acid metabolism and redox balance, especially biosynthesis of alanine and glutathione to exert its antioxidant effect.ROYAL SOC CHEMISTRY, 2015, METALLOMICS, 7(2) (2), 322 - 332, English[Refereed]Scientific journal
- Lignocellulosic biomass is a potential substrate for ethanol production. However, pretreatment of lignocellulosic materials produces inhibitory compounds such as acetic acid, which negatively affect ethanol production by Saccharomyces cerevisiae. Supplementation of the medium with three metal ions (Zn2+, Mg2+, and Ca2+) increased the tolerance of S. cerevisiae toward acetic acid compared to the absence of the ions. Ethanol production from xylose was most improved (by 34%) when the medium was supplemented with 2 mM Ca2+, followed by supplementation with 3.5 mM Mg2+ (29% improvement), and 180 mu M Zn2+ (26% improvement). Higher ethanol production was linked to high cell viability in the presence of metal ions. Comparative transcriptomics between the supplemented cultures and the control suggested that improved cell viability resulted from the induction of genes controlling the cell wall and membrane. Only one gene, FIT2, was found to be up-regulated in common between the three metal ions. Also up-regulation of HXT1 and TKL1 might enhance xylose consumption in the presence of acetic acid. Thus, the addition of ionic nutrients is a simple and cost-effective method to improve the acetic acid tolerance of S. cerevisiae.WILEY-V C H VERLAG GMBH, Dec. 2014, BIOTECHNOLOGY JOURNAL, 9(12) (12), 1519 - 1525, English[Refereed]Scientific journal
- Although the potential for biofuel production from microalgae via photosynthesis has been intensively investigated, information on the selection of a suitable operation strategy for microalgae-based biofuel production is lacking. Many published reports describe competitive strains and optimal culture conditions for use in biofuel production; however, the major impediment to further improvements is the absence of effective engineering strategies for microalgae cultivation and biofuel production. This comprehensive review discusses recent advances in understanding the effects of major environmental stresses and the characteristics of various engineering operation strategies on the production of biofuels (mainly biodiesel and bioethanol) using microalgae. The performances of microalgae-based biofuel-producing systems under various environmental stresses (i.e., irradiance, temperature, pH, nitrogen depletion, and salinity) and cultivation strategies (i.e., fed-batch, semi-continuous, continuous, two-stage, and salinity-gradient) are compared. The reasons for variations in performance and the underlying theories of the various production strategies are also critically discussed. The aim of this review is to provide useful information to facilitate development of innovative and feasible operation technologies for effectively increasing the commercial viability of microalgae-based biofuel production. (c) 2014 Elsevier Inc. All rights reserved.PERGAMON-ELSEVIER SCIENCE LTD, Dec. 2014, BIOTECHNOLOGY ADVANCES, 32(8) (8), 1448 - 1459, English[Refereed]Scientific journal
- Dec. 2014, Biotechnology for Biofuels, 7, 493, EnglishOverexpression of flv3 improves photosynthesis in the cyanobacterium Synechocystis sp. PCC6803 by enhancement of alternative electron flow[Refereed]Scientific journal
- Dec. 2014, Microbial Cell Factories, 13(1) (1), 173, EnglishDevelopment of bio-based fine chemical production through synthetic bioengineering[Refereed]Scientific journal
- Oligomeric sugars in the liquid fraction of hot water-pretreated rice straw are more amenable to membrane process than monomeric sugars, as lower pressure is required. Following membrane process was employed: nanofiltration (NF) concentration -> (dilution -> NF concentration) x 2 times -> enzymatic hydrolysis (EH) -> ultrafiltration (UF) permeation [Implication: NF for recovery of oligomeric sugars, dilution and NF for removal of low molecular weight fermentation inhibitors, UF for removal of high molecular weight fermentation inhibitors and recovery of monomeric sugars after EH]. This process provided the liquid fraction containing 111.4 g L-1 of sugars, corresponding to 681.0 mM as monomeric sugars, from the original liquid fraction (181.1 mM monomeric sugars). Concentrations of low molecular weight fermentation inhibitors, acetic and formic acids, were decreased to 24% and 48%, respectively. Xylose-fermenting recombinant Saccharomyces cerevisiae produced 34.5 +/- 2.2 g L-1 ethanol from the 0.8 times liquid fraction (76% of theoretical yield). (C) 2014 Published by Elsevier Ltd.ELSEVIER SCI LTD, Oct. 2014, BIORESOURCE TECHNOLOGY, 169, 380 - 386, English[Refereed]Scientific journal
- Oct. 2014, Microbial Cell Factories, 13(1) (1), 145, EnglishDevelopment of a GIN11/FRT-based multiple-gene integration technique affording inhibitor-tolerant, hemicellulolytic, xylose-utilizing abilities to industrial Saccharomyces cerevisiae strains for ethanol production from undetoxified lignocellulosic hemicel[Refereed]Scientific journal
- The toxic fermentation inhibitors in lignocellulosic hydrolysates raise serious problems for the microbial production of fuels and chemicals. Furfural is considered to be one of the most toxic compounds among these inhibitors. Here, we describe the detoxification of furfural in Corynebacterium glutamicum ATCC13032 under both aerobic and anaerobic conditions. Under aerobic culture conditions, furfuryl alcohol and 2-furoic acid were produced as detoxification products of furfural. The ratio of the products varied depending on the initial furfural concentration. Neither furfuryl alcohol nor 2-furoic acid showed any toxic effect on cell growth, and both compounds were determined to be the end products of furfural degradation. Interestingly, unlike under aerobic conditions, most of the furfural was converted to furfuryl alcohol under anaerobic conditions, without affecting the glucose consumption rate. Both the NADH/NAD(+) and NADPH/NADP(+) ratio decreased in the accordance with furfural concentration under both aerobic and anaerobic conditions. These results indicate the presence of a single or multiple endogenous enzymes with broad and high affinity for furfural and co-factors in C. glutamicum ATCC13032.SPRINGER, Oct. 2014, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 98(20) (20), 8675 - 8683, English[Refereed]Scientific journal
- Fuel ethanol produced from lignocettulose by the yeast Saccharomyces cerevisiae becomes an increasingly important alternative to fossil fuels. Selection of S. cerevisiae strains, which can effectively produce ethanol from xylose is crucial to improve the fuel yield from lignocellulose. In the present study, a universal calibration system was developed by the combination of time series, fermentation near infrared (NIR) spectral data analysis and reference high-performance analysis of a single yeast strain which enabled the evaluation of the ethanol production ability of a wide variety of xylose-fermenting yeast strains. Subtraction of xylose and ethanol concentrations at 0 h for each clone, as well as the respective spectra, reduced subtle errors of the fermentation components naturally occurring in multiple experiments to clearly visualize the difference of fermentation ability between strains. Also, NI R spectra showed specific peaks in difference spectra calculated from the subtraction treatment. A robust univariate linear regression model led to high prediction accuracy of xylose consumption (R-2 > 0.99) and ethanol production (R-2 > 0.98) for a wide variety of yeast strains when using only the distinct spectral pattern of a single-strain. Here, a novel method for screening of high-performing yeast strains has been developed requiring only a simple single-strain calibration model. The advantages of NIR spectroscopy such as rapid and convenient experimental preparation using electromagnetic waves in the region, which provide deep penetration into an aqueous sample, are successfully exploited in the proposed screening method.ROYAL SOC CHEMISTRY, Sep. 2014, ANALYTICAL METHODS, 6(17) (17), 6628 - 6634, English[Refereed]Scientific journal
- Jul. 2014, 日本エネルギー学会誌, 93(7) (7), 580 - 585, Japaneseアーミング酵母による統合型バイオプロセスの開発Scientific journal
- Simultaneous saccharification and fermentation (SSF) of renewable cellulose for the production of 3-phenyllactic acid (PhLA) by recombinant Escherichia coli was investigated. Kraft pulp recovered from biomass fractionation processes was used as a model cellulosic feedstock and was hydrolyzed using 10-50 filter paper unit (FPU) g(-1) kraft pulp of a commercial cellulase mixture, which increased the glucose yield from 21% to 72% in an enzyme dose-dependent manner. PhLA fermentation of the hydrolyzed kraft pulp by a recombinant E. coli strain expressing phenylpyruvate reductase from Wickerhamia fluorescens TK1 produced 1.9 mM PhLA. The PhLA yield obtained using separate hydrolysis and fermentation was enhanced from 5.8% to 42% by process integration into SSF of kraft pulp (20 g L-1) in a complex medium (pH 7.0) at 37 degrees C The PhLA yield was negatively correlated with the initial glucose concentration, with a five-fold higher PhLA yield observed in culture medium containing log L-1 glucose compared to 100 g L-1. Taken together, these results suggest that the PhLA yield from cellulose in kraft pulp can be improved by SSF under glucose-limited conditions. (C) 2014 Elsevier B.V. All rights reserved.ELSEVIER SCIENCE BV, Jul. 2014, BIOCHEMICAL ENGINEERING JOURNAL, 88, 188 - 194, English[Refereed]Scientific journal
- Jun. 2014, Biotechnology for Biofuels, 7, 97, EnglishOptimizing biodiesel production in marine Chlamydomonas sp.JSC4 through metabolic profiling and an innovative salinity-gradient strategy[Refereed]Scientific journal
- Jun. 2014, Biotechnology for Biofuels, 7, 88, EnglishGlycogen production for biofuels by the euryhaline cyanobacteria Synechococcus sp. strain PCC 7002 from an oceanic environment.[Refereed]Scientific journal
- Numerous environmental and social benefits could result from the replacement of petroleum-based transport fuels with bioethanol converted from renewable biomass. One of the key elements for the development of environmentally benign ethanol production is the construction of biomass-hydrolyzing yeast strains. The commonly used yeast Saccharomyces cerevisiae is a superior ethanol producer with demonstrated fast sugar consumption, high ethanol yield from glucose, and high resistance to ethanol. Hence, many researchers have targeted the heterologous expression of biomass-degrading enzymes in yeast to utilize glucose released from biomass for the production of ethanol by itself. In particular, cell-surface engineering is a powerful tool for yeast engineering. The display of amylolytic and cellulolytic enzymes on the yeast cell surface has accomplished direct ethanol production from starchy and cellulosic biomass. Moreover, reutilization of the cell surface-engineered yeast has the advantage of reducing enzyme cost, enabling reuse of enzymes on the cell surface by collecting the cells. For the efficient production of ethanol from biomass, improved assimilation of a wide variety of substrates could be achieved by overexpressing or deleting genes encoding traits responsible for yeast fermentability. Establishing economically feasible fermentation processes requires a marked increase in ethanol product titers due to the high energy demands of product recovery steps, as well as the high capital and production costs associated with bioethanol production equipment. A combination of biomass-degrading enzyme capacity and metabolic engineering in yeast strains could be an effective approach to developing cells with novel fermentation ability for industrial applications, and development of functional consolidated bioprocessing.Wiley Blackwell, Apr. 2014, Bioprocessing of Renewable Resources to Commodity Bioproducts, 201 - 226, English[Refereed]In book
- Mar. 2014, オレオサイエンス, 14(3) (3), 95 - 101, Japaneseリグノセルロースからの高効率エタノール生産に向けた新規代謝改変酵母の創製Scientific journal
- Mar. 2014, AMB Express, 4, 17, EnglishIncreased biomass production and glycogen accumulation in apcE gene deleted Synechocystis sp. PCC 6803[Refereed]Scientific journal
- Lignocellulosic biomass dedicated to bioethanol production usually contains pentoses and inhibitory compounds such as furfural that are not well tolerated by Saccharomyces cerevisiae. Thus, S. cerevisiae strains with the capability of utilizing both glucose and xylose in the presence of inhibitors such as furfural are very important in industrial ethanol production. Under the synergistic conditions of transaldolase (TAL) and alcohol dehydrogenase (ADH) overexpression, S. cerevisiae MT8-1X/TAL ADH was able to produce 1.3-fold and 2.3-fold more ethanol in the presence of 70 mM furfural than a TAL-expressing strain and a control strain, respectively. We also tested the strains' ability by mimicking industrial ethanol production from hemicellulosic hydrolysate containing fermentation inhibitors, and ethanol production was further improved by 16% when using MT8-1X/TAL-ADH compared to the control strain. Transcript analysis further revealed that besides the pentose phosphate pathway genes TKL1 and TAL1, ADH7 was also upregulated in response to furfural stress, which resulted in higher ethanol production compared to the TAL-expressing strain. The improved capability of our modified strain was based on its capacity to more quickly reduce furfural in situ resulting in higher ethanol production. The co-expression of TAL/ADH genes is one crucial strategy to fully utilize undetoxified lignocellulosic hydrolysate, leading to cost-competitive ethanol production. (C) 2013, The Society for Biotechnology, Japan. All rights reserved.SOC BIOSCIENCE BIOENGINEERING JAPAN, Feb. 2014, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 117(2) (2), 165 - 169, English[Refereed]Scientific journal
- Jan. 2014, FEBS LETTERS, 588(3) (3), 466 - 471[Refereed]
- Rre37 (sll1330) in a cyanobacterium Synechocystis sp. PCC 6803 acts as a regulatory protein for sugar catabolic genes during nitrogen starvation. Low glycogen accumulation in Delta rre37 was due to low expression of glycogen anabolic genes. In addition to low 2-oxoglutarate accumulation, normal upregulated expression of genes encoding glutamate synthases (gltD and gltB) as well as accumulation of metabolites in glycolysis (fructose-6-phosphate, fructose-1,6-bisphosphate, and glyceraldehyde-3-phosphate) and tricarboxylic acid (TCA) cycle (oxaloacetate, fumarate, succinate, and aconitate) were abolished by rre37 knockout. Rre37 regulates 2-oxoglutarate accumulation, glycogen accumulation through expression of glycogen anabolic genes, and TCA cycle metabolites accumulation. (C) 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.ELSEVIER SCIENCE BV, Jan. 2014, FEBS LETTERS, 588(3) (3), 466 - 471, English[Refereed]Scientific journal
- Jan. 2014, Biotechnology for Biofuels, 7(1) (1), 8, EnglishEfficient yeast cell surface display of exo-and endo-cellulase using the SED1 anchoring region and its original promoter[Refereed]Scientific journal
- Biodiesel production from microalgae has become a popular research topic. In this study, Chlamydomonas sp. JSC4 isolated from the southern coast of Taiwan was selected for a detailed study on cell growth and lipid accumulation under marine salinity (3.5% sea salt). Proper CO2 was supplied as the improvement of lipid productivity. Under the optimal condition, the highest lipid productivity was 169.1 mg/L/d, which was significantly higher than those reported in current studies for marine green algae. To date, only very few studies have reported a marine algae strain with both high cell growth and lipid productivity. This study demonstrated that a newly isolated marine green alga Chlamydomonas sp. JSC4 would be a feasible oil producer due to its high biomass production and lipid productivity under marine salinity. (C) 2013 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, Jan. 2014, BIORESOURCE TECHNOLOGY, 152, 247 - 252, English[Refereed]Scientific journal
- We tested the hypothesis that inducing photosynthesis in cyanobacteria requires respiration. A mutant deficient in glycogen phosphorylase (ΔGlgP) was prepared in Synechocystis sp. PCC 6803 to suppress respiration. The accumulated glycogen in ΔGlgP was 250-450% of that accumulated in wild type (WT). The rate of dark respiration in ΔGlgP was 25% of that inWT. In the dark, P700+ reduction was suppressed in ΔGlgP, and the rate corresponded to that in (2,5-dibromo-3-methyl-6-isopropyl- p -benzoquinone)-treated WT, supporting a lower respiration rate in ΔGlgP. Photosynthetic O2-evolution rate reached a steady-state value much slower in ΔGlgP than in WT. This retardation was solved by addition of D-glucose. Furthermore, we found that the contents of Calvin cycle intermediates in ΔGlgP were lower than those in WT under dark conditions. These observations indicated that respiration provided the carbon source for regeneration of ribulose 1,5-bisphosphate in order to drive the rapid start of photosynthesis.Japan Society for Bioscience Biotechnology and Agrochemistry, 2014, Bioscience, Biotechnology and Biochemistry, 78(12) (12), 1997 - 2007, English[Refereed]Scientific journal
- We produced organic acids, including lactate and succinate, directly from soluble starch under anaerobic conditions using high cell-density cultures of Corynebacterium glutamicum displaying α-amylase (AmyA) from Streptococcus bovis 148 on the cell surface. Notably, reactions performed under anaerobic conditions at 35 and 40°C, which are higher than the optimal growth temperature of 30°C, showed 32% and 19%, respectively, higher productivity of the organic acids lactate, succinate, and acetate compared to that at 30°C. However, α-amylase was not stably anchored and released into the medium from the cell surface during reactions at these higher temperatures, as demonstrated by the 61% and 85% decreases in activity, respectively, from baseline, compared to the only 8% decrease at 30°C. The AmyA-displaying C. glutamicum cells retained their starch-degrading capacity during five 10 h reaction cycles at 30°C, producing 107.8 g/l of total organic acids, including 88.9 g/l lactate and 14.0 g/l succinate. The applicability of cell surface-engineering technology for the production of organic acids from biomass by high cell-density cultures of C. glutamicum under anaerobic conditions was demonstrated.Dec. 2013, AMB Express, 3(1) (1), 72 - 72, English, International magazine[Refereed]Scientific journal
- Concentrating sugars using membrane separation, followed by ethanol fermentation by recombinant xylose-assimilating Saccharomyces cerevisiae, is an attractive technology. Three nanofiltration membranes (NTR-729HF, NTR-7250, and ESNA3) were effective in concentrating glucose, fructose, and sucrose from dilute molasses solution and no permeation of sucrose. The separation factors of acetate, formate, furfural, and 5-hydroxymethyl furfural, which were produced by dilute acid pretreatment of rice straw, over glucose after passage through these three membranes were 3.37-11.22, 4.71-20.27, 4.32-16.45, and 4.05-16.84, respectively, at pH 5.0, an applied pressure of 1.5 or 2.0 MPa, and 25 degrees C. The separation factors of these fermentation inhibitors over xylose were infinite, as there was no permeation of xylose. Ethanol production from approximately two-times concentrated liquid hydrolysate using recombinant S. cerevisiae was double (5.34-6.44 g L-1) that compared with fermentation of liquid hydrolysate before membrane separation (2.75 g L-1). (C) 2013 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, Nov. 2013, BIORESOURCE TECHNOLOGY, 147, 84 - 88, English[Refereed]Scientific journal
- With the exhaustion of fossil fuels and with the environmental issues they pose, utilization of abundant lignocellulosic biomass as a feedstock for biofuels and bio-based chemicals has recently become an attractive option. Lignocellulosic biomass is primarily composed of cellulose, hemicellulose, and lignin and has a very rigid and complex structure. It is accordingly much more expensive to process than starchy grains because of the need for extensive pretreatment and relatively large amounts of cellulases for efficient hydrolysis. Efficient and cost-effective methods for the production of biofuels and chemicals from lignocellulose are required. A consolidated bioprocess (CBP), which integrates all biological steps consisting of enzyme production, saccharification, and fermentation, is considered a promising strategy for reducing production costs. Establishing an efficient CBP using lignocellulosic biomass requires both lignocellulose degradation into glucose and efficient production of biofuels or chemicals from glucose. With this aim, many researchers are attempting to endow selected microorganisms with lignocellulose-assimilating ability. In this review, we focus on studies aimed at conferring lignocellulose-assimilating ability not only to yeast strains but also to bacterial strains by recombinant technology. Recent developments in improvement of enzyme productivity by microorganisms and in improvement of the specific activity of cellulase are emphasized. (C) 2013 Elsevier Inc. All rights reserved.PERGAMON-ELSEVIER SCIENCE LTD, Nov. 2013, BIOTECHNOLOGY ADVANCES, 31(6) (6), 754 - 763, English[Refereed]Scientific journal
- Agricultural residues comprising lignocellulosic materials are excellent sources of pentose sugar, which can be converted to ethanol as fuel. Ethanol production via consolidated bioprocessing requires a suitable microorganism to withstand the harsh fermentation environment of high temperature, high ethanol concentration, and exposure to inhibitors. We genetically enhanced an industrial Saccharomyces cerevisiae strain, sun049, enabling it to uptake xylose as the sole carbon source at high fermentation temperature. This strain was able to produce 13.9 g/l ethanol from 50 g/l xylose at 38 A degrees C. To better understand the xylose consumption ability during long-term, high-temperature conditions, we compared by transcriptomics two fermentation conditions: high temperature (38 A degrees C) and control temperature (30 A degrees C) during the first 12 h of fermentation. This is the first long-term, time-based transcriptomics approach, and it allowed us to discover the role of heat-responsive genes when xylose is the sole carbon source. The results suggest that genes related to amino acid, cell wall, and ribosomal protein synthesis are down-regulated under heat stress. To allow cell stability and continuous xylose uptake in order to produce ethanol, hexose transporter HXT5, heat shock proteins, ubiquitin proteins, and proteolysis were all induced at high temperature. We also speculate that the strong relationship between high temperature and increased xylitol accumulation represents the cell's mechanism to protect itself from heat degradation.SPRINGER HEIDELBERG, Sep. 2013, JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 40(9) (9), 1039 - 1050, English[Refereed]Scientific journal
- Engineering strategies were applied to improve the cell growth, CO2 fixation ability, and carbohydrate productivity of a Scenedesmus obliquus CNW-N isolate. The resulting carbohydrate-rich microalgal biomass was subsequently utilized as feedstock for ethanol fermentation. The microalga was cultivated with 2.5% CO2 in a photobioreactor on different operation modes. Semi-batch operations with 50% replacement of culture medium resulted in the highest CO2 fixation rate (1546.7 mg L-1 d(-1)), carbohydrate productivity (467.6 mg L-1 d(-1)), and bioethanol yield (0.202 g/g biomass). This performance is better than most reported values in the literature. The microalgal biomass can accumulate nearly 50% carbohydrates, as glucose accounted for nearly 80% of the total carbohydrate content. This glucose-predominant carbohydrate composition of the microalga is well suited for fermentative bioethanol production. Therefore, using the proposed carbohydrate-rich microalgal biomass both as the carbon sink and as the feedstock provides a feasible alternative to current carbon-reduction and bioethanol-production strategies. (C) 2013 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, Sep. 2013, BIORESOURCE TECHNOLOGY, 143, 163 - 171, English[Refereed]Scientific journal
- Cocktail delta-integration was applied to improve ethanol production from xylose in Saccharomyces cerevisiae. Two hundred of recombinant S. cerevisiae strains possessing various copies of XYL1, XYL2, and XKS1 genes were constructed by cocktail delta-integration. Efficient strains with efficient ethanol production from xylose were successfully obtained by the fermentation test. (C) 2013, The Society for Biotechnology, Japan. All rights reserved.SOC BIOSCIENCE BIOENGINEERING JAPAN, Sep. 2013, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 116(3) (3), 333 - 336, English[Refereed]Scientific journal
- Cyanobacterial glycogen has gained interest as a valuable biomass feedstock for biofuel production. However, an ideal method for native glycogen quantification has not been developed. Here, we have proposed a simple methodology that enables the quantitative determination of cyanobacterial glycogen concentration with high repeatability using aqueous size-exclusion chromatography with a differential refractive index detector (SEC/RID). Our SEC/RID system also allows size distributions for native glycogen based on hydrodynamic volumes (Vh), which is proportional to the product of the molecular mass (M) and intrinsic viscosity [η], obtained by universal calibration using linear homopolymers of known M with Mark-Houwink-Sakurada parameters. The universal calibration curve achieved a broad linear range (Vh parameter [η]M=2×102-8×108mLg-1) with a high correlation coefficient (R2=0.9942), because the developed system is equipped with an OHpak SB-806M HQ aqueous column containing four types of polyhydroxy methacrylate-based particles with different particle and pore sizes. Based on the SEC/RID system, response of molecular size distribution of glycogen in microalgae to the cultivation condition was first observed. Our established SEC/RID method has several advantages over conventional techniques, including the simultaneous quantitative and size distribution analyses of glycogen, and represents a potentially useful tool to elucidate the relationship between structural properties and the roles of glycogen in metabolism. © 2013 Elsevier B.V.Jul. 2013, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 930, 90 - 97, English[Refereed]Scientific journal
- Jun. 2013, Journal of Experimental Botany, 64(10) (10), 2943 - 2954, EnglishDynamic metabolic profiling of cyanobacterial glycogen biosynthesis under conditions of nitrate depletion[Refereed]Scientific journal
- Simple pharmacological studies on inositol stereoisomers are presented in this study. Male ICR mice were orally administered 1 g/kg BW of three inositol stereoisomers, myo-inositol (MI), d-chiro-inositol (DCI), and scyllo-inositol (SI), and blood plasma samples and skeletal muscle fractions were prepared after an hour. The plasma samples were subjected to gas chromatography-coupled time-of-flight mass spectrometry (GC-TOF-MS) analysis. None of the three stereoisomers was seen in untreated samples, but substantial amounts ranging from 2.5 to 6.5 mM were detected only after administration, indicating that orally administered inositol stereoisomers were readily absorbed and their levels elevated in the bloodstream. In addition, plasma of SI-administered animals contained substantial MI, suggesting a possible metabolic conversion of SI to MI. In the skeletal muscle fractions, glucose transporter type 4 (GLUT4) content in the plasma membrane increased, indicating that inositol stereoisomers stimulated GLUT4 translocation. © 2013 American Chemical Society.May 2013, Journal of Agricultural and Food Chemistry, 61(20) (20), 4850 - 4854, English[Refereed]Scientific journal
- May 2013, Energy & Environmental Science, 6(6) (6), 1844 - 1849, EnglishDirect conversion of Spirulina to ethanol without pretreatment or enzymatic hydrolysis processes[Refereed]Scientific journal
- The aim of this study is to develop a scheme of cell recycle batch fermentation (CRBF) of high-solid lignocellulosic materials. Two-phase separation consisting of rough removal of lignocellulosic residues by low-speed centrifugation and solid-liquid separation enabled effective collection of Saccharomyces cerevisiae cells with decreased lignin and ash. Five consecutive batch fermentation of 200 g/L rice straw hydrothermally pretreated led to an average ethanol titer of 34.5 g/L. Moreover, the display of cellulases on the recombinant yeast cell surface increased ethanol titer to 42.2 g/L. After, five-cycle fermentation, only 3.3 g/L sugar was retained in the fermentation medium, because cellulase displayed on the cell surface hydrolyzed cellulose that was not hydrolyzed by commercial cellulases or free secreted cellulases. Fermentation ability of the recombinant strain was successfully kept during a five-cycle repeated batch fermentation with 86.3% of theoretical yield based on starting biomass. (C) 2012 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, May 2013, BIORESOURCE TECHNOLOGY, 135, 403 - 409, English[Refereed]Scientific journal
- This study aimed to evaluate the potential of using a carbohydrate-rich microalga Chlorella vulgaris FSP-E as feedstock for bioethanol production via various hydrolysis strategies and fermentation processes. Enzymatic hydrolysis of C. vulgaris FSP-E biomass (containing 51% carbohydrate per dry weight) gave a glucose yield of 90.4% (or 0.461 g (g biomass)(-1)). The SHF and SSF processes converted the enzymatic microalgae hydrolysate into ethanol with a 79.9% and 92.3% theoretical yield, respectively. Dilute acidic hydrolysis with 1% sulfuric acid was also very effective in saccharifying C vulgaris FSP-E biomass, achieving a glucose yield of nearly 93.6% from the microalgal carbohydrates at a starting biomass concentration of 50 g L-1. Using the acidic hydrolysate of C vulgaris FSP-E biomass as feedstock, the SHF process produced ethanol at a concentration of 11.7 g L-1 and an 87.6% theoretical yield. These findings indicate the feasibility of using carbohydrate-producing microalgae as feedstock for fermentative bioethanol production. (C) 2012 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, May 2013, BIORESOURCE TECHNOLOGY, 135, 191 - 198, English[Refereed]Scientific journal
- The biorefinery manufacturing process for producing chemicals and liquid fuels from biomass is a promising approach for securing energy and resources. To establish cost-effective fermentation of lignocellulosic biomass, the consolidation of sacccharification and fermentation processes is a desirable strategy, but requires the development of microorganisms capable of cellulose/hemicellulose hydrolysis and target chemical production. Such an endeavor requires a large number of prerequisites to be realized, including engineering microbial strains with high cellulolytic activity, high product yield, productivities, and titers, ability to use many carbon sources, and resistance to toxic compounds released during the pretreatment of lignocellulosic biomass. Researchers have focused on either engineering naturally cellulolytic microorganisms to improve product-related properties or modifying non-cellulolytic organisms with high product yields to become cellulolytic. This article reviews recent advances in the development of microorganisms for the production of renewable chemicals and advanced biofuels, as well as ethanol, from lignocellulosic materials through consolidated bioprocessing. (C) 2012 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, May 2013, BIORESOURCE TECHNOLOGY, 135, 513 - 522, English[Refereed]Scientific journal
- Enzymatic hydrolysis of cellulosic material is an essential step in the bioethanol production process. However, complete cellulose hydrolysis by cellulase is difficult due to the irreversible adsorption of cellulase onto cellulose. Thus, part of the cellulose remains in crystalline form after hydrolysis. In this study, after 96-h hydrolysis of Avicel crystalline cellulose, 47.1 % of the cellulase was adsorbed on the cellulose surface with 10.8 % crystalline cellulose remaining. In simultaneous saccharification and fermentation of 100 g/L Avicel with 1.0 filter paper unit/mL cellulase, a wild-type yeast strain produced 44.7 g/L ethanol after 96 h. The yield of ethanol was 79.7 % of the theoretical yield. On the other hand, a recombinant yeast strain displaying various cellulases, such as β-glucosidase, cellobiohydrolase, and endoglucanase, produced 48.9 g/L ethanol, which corresponds to 87.3 % of the theoretical yield. Higher ethanol production appears to be attributable to higher efficiency of cellulase displayed on the cell surface. These results suggest that cellulases displayed on the yeast cell surface improve hydrolysis of Avicel crystalline cellulose. Indeed, after the 96-h simultaneous saccharification and fermentation using the cellulase-displaying yeast, the amount of residual cellulose was 1.5 g/L, one quarter of the cellulose remaining using the wild-type strain, a result of the alleviation of irreversible adsorption of cellulases on the crystalline cellulose. © 2012 Springer-Verlag Berlin Heidelberg.Mar. 2013, Applied Microbiology and Biotechnology, 97(5) (5), 2231 - 2237, English[Refereed]Scientific journal
- Several alcohol dehydrogenase (ADH)-related genes have been identified as enzymes for reducing levels of toxic compounds, such as, furfural and/or 5-hydroxymethylfurfural (5-HMF), in hydrolysates of pretreated lignocelluloses. To date, overexpression of these ADH genes in yeast cells have aided ethanol production from glucose or glucose/xylose mixture in the presence of furfural or 5-HMF. However, the effects of these ADH isozymes on ethanol production from xylose as a sole carbon source remain uncertain. We showed that overexpression of mutant NADH-dependent ADH1 derived from TMB3000 strain in the recombinant Saccharomyces cerevisiae, into which xylose reductase (XR) and xylitol dehydrogenase (XDH) pathway of Pichia stipitis has been introduced, improved ethanol production from xylose as a sole carbon source in the presence of 5-HMF. Enhanced furan-reducing activity is able to regenerate NAD+ to relieve redox imbalance, resulting in increased ethanol yield arising from decreased xylitol accumulation. In addition, we found that overexpression of wild-type ADH1 prevented the more severe inhibitory effects of furfural in xylose fermentation as well as overexpression of TMB3000-derived mutant. After 120 h of fermentation, the recombinant strains overexpressing wild-type and mutant ADH1 completely consumed 50 g/L xylose in the presence of 40 mM furfural and most efficiently produced ethanol (15.70 g/L and 15.24 g/L) when compared with any other test conditions. This is the first report describing the improvement of ethanol production from xylose as the sole carbon source in the presence of furan derivatives with xylose-utilizing recombinant yeast strains via the overexpression of ADH-related genes. © 2012 Springer-Verlag.Mar. 2013, Applied Microbiology and Biotechnology, 97(6) (6), 2597 - 2607, English[Refereed]Scientific journal
- Three enzymes responsible for the transhydrogenase-like shunt, including malic enzyme (encoded by MAE1), malate dehydrogenase (MDH2), and pyruvate carboxylase (PYC2), were overexpressed to regulate the redox state in xylose-fermenting recombinant Saccharomyces cerevisiae. The YPH499XU/MAE1 strain was constructed by overexpressing native Mae1p in the YPH499XU strain expressing xylose reductase and xylitol dehydrogenase from Scheffersomyces stipitis, and native xylulokinase. Analysis of the xylose fermentation profile under semi-anaerobic conditions revealed that the ethanol yield in the YPH499XU/MAE1 strain (0.38 +/- 0.01 g g(-1) xylose consumed) was improved from that of the control strain (0.31 +/- 0.01 g g(-1) xylose consumed). Reduced xylitol production was also observed in YPH499XU/MAE1, suggesting that the redox balance was altered by Mae1p overexpression. Analysis of intracellular metabolites showed that the redox imbalance during xylose fermentation was partly relieved in the transformant. The specific ethanol production rate in the YPH499XU/MAE1-MDH2 strain was 1.25-fold higher than that of YPH499XU/MAE1 due to the additional overexpression of Mdh2p, whereas the ethanol yield was identical to that of YPH499XU/MAE1. The specific xylose consumption rate was drastically increased in the YPH499XU/MAE1-MDH2-PYC2 strain. However, poor ethanol yield as well as increased production of xylitol was observed. These results demonstrate that the transhydrogenase function implemented in S. cerevisiae can regulate the redox state of yeast cells.SPRINGER, Feb. 2013, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 97(4) (4), 1669 - 1678, English[Refereed]Scientific journal
- A perfluoropolymer (PFP) membrane has been prepared for use in vapor permeation to separate aqueous ethanol mixtures produced from rice straw with xylose-assimilating recombinant Saccharomyces cerevisiae. PFP membranes commonly have been used for dehydration process and possess good selectivity and high permeances. The effects of by-products during dilute acid pretreatment, addition of yeast extract, and ethanol fermentation on PFP membrane performance were investigated. While feeding mixtures of ethanol (90wt%) in water, to which individual by-products (0.1-2g/L) were added, the PFP membrane demonstrated no clear change in permeation rate (439-507gm-2h-1) or separation factor (14.9-23.5) from 2 to 4h of the process. The PFP membrane also showed no clear change in permeation rate (751-859gm-2h-1) or separation factor (12.5-13.8) while feeding the mixture (final ethanol conc.: 61wt%) of ethanol and distillation of the fermentation broth using a suspended fraction of dilute acid-pretreated rice straw for 20h. These results suggest that the PFP membrane can tolerate actual distillation liquids from ethanol fermentation broth obtained from lignocellulosic biomass pretreated with dilute acid. © 2012.Jan. 2013, Biochemical Engineering Journal, 70, 135 - 139, English[Refereed]Scientific journal
- Production of ethanol from xylose at high temperature would be an economical approach since it reduces risk of contamination and allows both the saccharification and fermentation steps in SSF to be running at elevated temperature. Eight recombinant xylose-utilizing Saccharomyces cerevisiae strains developed from industrial strains were constructed and subjected to high-temperature fermentation at 38 °C. The best performing strain was sun049T, which produced up to 15.2. g/L ethanol (63% of the theoretical production), followed by sun048T and sun588T, both with 14.1. g/L ethanol produced. Via transcriptomic analysis, expression profiling of the top three best ethanol producing strains compared to a negative control strain, sun473T, led to the discovery of genes in common that were regulated in the same direction. Identification of the 20 most highly up-regulated and the 20 most highly down-regulated genes indicated that the cells regulate their central metabolism and maintain the integrity of the cell walls in response to high temperature. We also speculate that cross-protection in the cells occurs, allowing them to maintain ethanol production at higher concentration under heat stress than the negative controls. This report provides further transcriptomics information in the interest of producing a robust microorganism for high-temperature ethanol production utilizing xylose. © 2012 Elsevier B.V.Jan. 2013, Journal of Biotechnology, 163(1) (1), 50 - 60, English[Refereed]Scientific journal
- Synthetic bioengineering is a strategy for developing useful microbial strains with innovative biological functions. Novel functions are designed and synthesized in host microbes with the aid of advanced technologies for computer simulations of cellular processes and the system-wide manipulation of host genomes. Here, we review the current status and future prospects of synthetic bioengineering in the yeast Saccharomyces cerevisiae for bio-refinery processes to produce various commodity chemicals from lignocellulosic biomass. Previous studies to improve assimilation of xylose and production of glutathione and butanol suggest a fixed pattern of problems that need to be solved, and as a crucial step, we now need to identify promising targets for further engineering of yeast metabolism. Metabolic simulation, transcriptomics, and metabolomics are useful emerging technologies for achieving this goal, making it possible to optimize metabolic pathways. Furthermore, novel genes responsible for target production can be found by analyzing large-scale data. Fine-tuning of enzyme activities is essential in the latter stage of strain development, but it requires detailed modeling of yeast metabolic functions. Recombinant technologies and genetic engineering are crucial for implementing metabolic designs into microbes. In addition to conventional gene manipulation techniques, advanced methods, such as multicistronic expression systems, marker-recycle gene deletion, protein engineering, cell surface display, genome editing, and synthesis of very long DNA fragments, will facilitate advances in synthetic bioengineering. (C) 2012 Elsevier B.V. All rights reserved.ELSEVIER SCIENCE BV, Jan. 2013, JOURNAL OF BIOTECHNOLOGY, 163(2) (2), 204 - 216, English[Refereed]Scientific journal
- In this study, three indigenous microalgae isolates were examined for their ability to produce carbohydrates. Among them, Chlorella vulgaris FSP-E displayed relatively high cell growth rate and carbohydrate content. The carbohydrate productivity of C. vulgaris FSP-E was further improved by using engineering strategies. The results show that using an appropriate light intensity and inoculum size could effectively promote cell growth and carbohydrate productivity. Nitrogen starvation triggered the accumulation of carbohydrates in the microalga, achieving a carbohydrate content of 51.3% after 4-day starvation. Under the optimal conditions, the highest biomass and carbohydrate productivity were 1.437 and 0.631gL-1d-1, respectively. This performance is better than that reported in most related studies. Since glucose accounted for nearly 93% of the carbohydrates accumulated in C. vulgaris FSP-E, the microalga is an excellent feedstock for bioethanol fermentation. © 2012 Elsevier Ltd.Elsevier Ltd, 2013, Bioresource Technology, 135, 157 - 165, English[Refereed]Scientific journal
- To build an energy and material secure future, a next generation of renewable fuels produced from lignocellulosic biomass is required. Although lignocellulosic biomass, which represents an abundant, inexpensive and renewable source for bioethanol production, is of great interest as a feedstock, the complicated ethanol production processes involved make the cost of producing bioethanol from it higher compared to corn starch and cane juice. Therefore, consolidated bioprocessing (CBP), which combines enzyme production, saccharification and fermentation in a single step, has gained increased recognition as a potential bioethanol production system. CBP requires a highly engineered microorganism developed for several different process-specific characteristics. The dominant strategy for engineering a CBP biocatalyst is to express multiple components of a cellulolytic system from either fungi or bacteria in the yeast Saccharomyces cerevisiae. The development of recombinant yeast strains displaying cellulases and hemicellulases on the cell surface represents significant progress toward realization of CBP. Regardless of the process used for biomass hydrolysis. CBP-enabling microorganisms encounter a variety of toxic compounds produced during biomass pretreatment that inhibit microbial growth and ethanol yield. Systems biology approaches including disruptome screening, transcriptomics, and metabolomics have been recently exploited to gain insight into the molecular and genetic traits involved in tolerance and adaptation to the fermentation inhibitors. In this review, we locus on recent advances in development of yeast strains with both the ability to directly convert lignocellulosic material to ethanol and tolerance in the harsh environments containing toxic compounds in the presence of ethanol. (c) 2011 Elsevier Inc. All rights reserved.PERGAMON-ELSEVIER SCIENCE LTD, Nov. 2012, BIOTECHNOLOGY ADVANCES, 30(6) (6), 1207 - 1218, English[Refereed]Scientific journal
- Oct. 2012, ペトロテック, 35(10) (10), 700 - 706, JapaneseバイオリファイナリーとバイオプラスチックScientific journal
- Consolidated bioprocessing (CBP), which integrates enzyme production, saccharification and fermentation into a single process, is a promising strategy for effective ethanol production from lignocellulosic materials because of the resulting reduction in utilities, the substrate and other raw materials and simplification of operation. CBP requires a highly engineered microbial strain capable of hydrolyzing biomass with enzymes produced on its own and producing high-titer ethanol. Recently, heterologous production of cellulolytic enzymes has been pursued with yeast hosts, which has realized direct conversion of cellulose to ethanol. Specifically, the development of cell surface engineering, which provides a display of cellulolytic enzymes on the yeast cell surface, facilitates effective biomass hydrolysis concomitantly with ethanol production. On the other hand, the difference in optimum temperature between saccharification and fermentation is a drawback of efficient ethanol production in the simultaneous saccharification and fermentation (SSF). The application of thermotolerant yeast strains engineered to the SSF process would overcome the drawback by performing hydrolysis and fermentation at elevated temperature. In this review, we focus on the recent advances in the application of thermotolerant yeast to CBP and SSF of lignocellulosic material to ethanol. The development of thermotolerant and ethanologenic yeast strains with the ability to hydrolyze lignocellulosic materials is emphasized for high-temperature CBP. (C) 2012 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, Sep. 2012, PROCESS BIOCHEMISTRY, 47(9) (9), 1287 - 1294, English[Refereed]Scientific journal
- To build an energy and material secure future, we must pioneer the next generation of renewable fuels and chemicals using environmentally benign production processes. Since biomass represents an abundant carbon-neutral renewable resource for the production of biofuels, numerous environmental and social benefits could result from the replacement of petroleum-based transport fuels with bioethanol converted from biomass. One of the key technologies for the development of biorefineries is cell surface engineering, which is a powerful tool for engineering and functionalizing many organisms. Using the technology, various kinds of functional proteins, such as enzymes, can be expressed on the cell surface without loss of cell activity. The display of amylolytic and cellulolytic enzymes on the surface of Saccharomyces cerevisiae has accomplished direct ethanol production from starchy and cellulosic biomass. Moreover, the display of hemicellulase on the surface of S. cerevisiae that has a xylose-assimilating metabolic pathway has enabled production of ethanol from hemicellulosic materials. Furthermore, reutilization of the cell-surface engineered yeast has an advantage in the reduction of enzyme cost, which enables reuse of enzymes on the cell surface by collecting the cells. Thus, cell surface engineering is a promising technology for the development of a consolidated bioprocess by integrating enzyme production, saccharification and fermentation. Regardless of the biomass hydrolysis, metabolic engineering of microorganisms is emphasized for the efficient production of ethanol from biomass. Specifically, lignocellulosic hydrolysates contain high concentrations of inhibitors that negatively affect metabolism and ethanol yields. To circumvent these difficulties, robust S. cerevisiae strains that efficiently ferment mixtures of hexose and pentose sugars in the presence of various chemical contexts for industrial ethanol production should be constructed through metabolic engineering approaches. A combination of a cell-surface displayed enzyme system and an intracellular metabolic engineering system is a very effective approach for developing cells with improved fermentation ability for industrial applications. The technology (synthetic bioengineering) will open up new applications of cell factories to industrially important processes.Springer Netherlands, Aug. 2012, Systems Metabolic Engineering, 9789400745346, 329 - 348, English[Refereed]In book
- Jul. 2012, 生物工学会誌, 90(7) (7), 386 - 391, Japanese合成生物工学によるバイオ燃料生産のための微生物細胞工場の創製Scientific journal
- To improve the ability of recombinant Saccharomyces cerevisiae strains to utilize the hemicellulose components of lignocellulosic feedstocks, the efficiency of xylose conversion to ethanol needs to be increased. In the present study, xylose-fermenting, haploid, yeast cells of the opposite mating type were hybridized to produce a diploid strain harboring two sets of xylose-assimilating genes encoding xylose reductase, xylitol dehydrogenase, and xylulokinase. The hybrid strain MN8140XX showed a 1.3- and 1.9-fold improvement in ethanol production compared to its parent strains MT8-1X405 and NBRC1440X, respectively. The rate of xylose consumption and ethanol production was also improved by the hybridization. This study revealed that the resulting improvements in fermentation ability arose due to chromosome doubling as well as the increase in the copy number of xylose assimilation genes. Moreover, compared to the parent strain, the MN8140XX strain exhibited higher ethanol production under elevated temperatures (38 A degrees C) and acidic conditions (pH 3.8). Thus, the simple hybridization technique facilitated an increase in the xylose fermentation activity.SPRINGER, Jun. 2012, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 94(6) (6), 1585 - 1592, English[Refereed]Scientific journal
- A method for a widely targeted analysis was developed for the metabolic profiling of yeast central metabolism. The widely targeted method consists of 2 analyses, namely, gas chromatography-quadrupole-mass spectrometry (GC-Q-MS) operated in selected ion monitoring mode with 25 m/z channels, and liquid chromatography triple-stage quadrupole (LC-QqQ)-MS operated in multiple reaction monitoring mode. This platform was set up to identify and quantify preselected 99 compounds, including sugars, sugar phosphates, organic adds, amino acids, and cofactors. The method showed good sensitivity and a wide dynamic range. For example, limits of detection for lactate and L-phenylalanine were 1.4 fmol and 2.0 fmol, respectively. The dynamic ranges for GC-Q-MS analysis and LC-QqQ-MS analysis were approximately 10(2)-10(5) and 10(3)-10(4), respectively. The metabolite profiles of 2 yeast strains, YPH499 and BY4741, under glucose-fermenting conditions were compared using the developed method. Although YPH499 and BY4741 were derived from an identical experimental strain, the profiling analysis successfully revealed a variation in metabolic phenotypes among experimental yeast strains demonstrating that the widely targeted method could be a robust and useful method for the investigation of metabolic phenotypes of Saccharomyces cerevisiae. (C) 2012, The Society for Biotechnology, Japan. All rights reserved.SOC BIOSCIENCE BIOENGINEERING JAPAN, May 2012, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 113(5) (5), 665 - 673, English[Refereed]Scientific journal
- For efficient bioethanol production from lignocellulosic biomass by Saccharomyces cerevisiae, it is necessary to improve cellular tolerance to toxic compounds released during the pretreatment of biomass. The gene encoding p-nitrophenylphosphatase, PHO13, was disrupted in a recombinant xylose-fermenting S. cerevisiae strain, which improved ethanol production from xylose in the presence of three major inhibitors, acetic and formic acids, and furfural. In medium supplemented with 30 mM acetic acid, the ethanol yield obtained by the Delta PHO13 mutant was 0.45 g-ethanol/g-xylose. Notably, the specific ethanol productivity of the mutant in the presence of 90 mM furfural was fourfold higher than that of the control strain. The PHO13-disrupted strain produced ethanol from rice straw hydrolysate obtained by liquid hot-water pretreatment with a greater than fourfold higher xylose consumption rate than the control. Together, our findings demonstrate that PHO13 deletion is a simple, but effective, approach for improving cellulosic bioethanol production by S. cerevisiae. (C) 2012 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, May 2012, BIORESOURCE TECHNOLOGY, 111, 161 - 166, English[Refereed]Scientific journal
- The cost of the lignocellulose-hydrolyzing enzymes used in the saccharification process of ethanol production from biomass accounts for a relatively high proportion of total processing costs. Cell surface engineering technology has facilitated a reduction in these costs by integrating saccharification and fermentation processes into a recombinant microbe strain expressing heterologous enzymes on the cell surface. We constructed a recombinant Saccharomyces cerevisiae that not only hydrolyzed hemicelluloses by codisplaying endoxylanase from Trichoderma reesei, beta-xylosidase from Aspergillus oryzae, and beta-glucosidase from Aspergillus aculeatus but that also assimilated xylose through the expression of xylose reductase and xylitol dehydrogenase from Pichia stipitis and xylulokinase from S. cerevisiae. The recombinant strain successfully produced ethanol from rice straw hydrolysate consisting of hemicellulosic material containing xylan, xylooligosaccharides, and cellooligosaccharides without requiring the addition of sugar-hydrolyzing enzymes or detoxication. The ethanol titer of the strain was 8.2 g/l after 72 h fermentation, which was approximately 2.5-fold higher than that of the control strain. The yield (grams of ethanol per gram of total sugars in rice straw hydrolysate consumed) was 0.41 g/g, which corresponded to 82% of the theoretical yield. The cell surface-engineered strain was thus highly effective for consolidating the process of ethanol production from hemicellulosic materials. (C) 2011 Elsevier B.V. All rights reserved.ELSEVIER SCIENCE BV, Apr. 2012, JOURNAL OF BIOTECHNOLOGY, 158(4) (4), 203 - 210, English[Refereed]Scientific journal
- Mar. 2012, Bioresource Technology, 108, 211-215, EnglishSynergistic enhancement of glycogen production in Arthrospira platensis by optimization of light intensity and nitrate supply[Refereed]Scientific journal
- Mar. 2012, Bioresource Technology, 108, 128-133, EnglishDisplay of cellulases on the cell surface of Saccharomyces cerevisiae affords high yield ethanol production from high-solid lignocellulosic biomass[Refereed]Scientific journal
- A major challenge associated with the fermentation of lignocellulose-derived hydrolysates is improved ethanol production in the presence of fermentation inhibitors, such as acetic and formic acids. Enhancement of transaldolase (TAL) and formate dehydrogenase (FDH) activities through metabolic engineering successfully conferred resistance to weak acids in a recombinant xylose-fermenting Saccharomyces cerevisiae strain. Moreover, hybridization of the metabolically engineered yeast strain improved ethanol production from xylose in the presence of both 30 mM acetate and 20 mM formate. Batch fermentation of lignocellulosic hydrolysate containing a mixture of glucose, fructose and xylose as carbon sources, as well as the fermentation inhibitors, acetate and formate, was performed for five cycles without any loss of fermentation capacity. Long-term stability of ethanol production in the fermentation phase was not only attributed to the coexpression of TAL and FDH genes, but also the hybridization of haploid strains. (C) 2011 Elsevier Ltd. All rights reserved.ELSEVIER SCI LTD, Sep. 2011, BIORESOURCE TECHNOLOGY, 102(17) (17), 7917 - 7924, English[Refereed]Scientific journal
- Aug. 2011, Bioscience, Biotechnology, and Biochemistry, 75(8) (8), 1603 - 1605, EnglishVariation in biomass property among rice diverse cultivars.[Refereed]Scientific journal
- Recently, genetic engineering efforts have been made to develop recombinant Saccharomyces cerevisiae strains able to utilize xylose, an inexpensive and abundant carbon source. However, their construction and selection processes are limited by the speed and expenses of the existing testing methods, thus a rapid and equally precise method will significantly increase the number of tested strains. Here, near infrared (NIR) spectroscopy is proposed as a successful alternative method for screening recombinant xylose-fermenting S. cerevisiae. Supernatant samples of fermentation solutions from one diploid and three haploid recombinant strains were collected along the fermentation process. NIR spectra of the diluted supernatant provided effective differentiation of strains consistent with their phenotypic and genotypic features. This result could be used as a feedback for multicomponent analysis, in order to develop regression model for quantification of consumed glucose and xylose, produced ethanol, glycerol, and xylitol. Robust partial least-squares regression models with high prediction accuracy that are effective with any strain were achieved for all components when the modeling was performed with combined data of all strains, achieving 0.21-1.49 g/L of standard error of prediction with calibration, prediction, limit of detection and limit of quantification in the range of 1.0-4.5 and 3.0-13.4 g/L, respectively.AMER CHEMICAL SOC, Jun. 2011, ANALYTICAL CHEMISTRY, 83(11) (11), 4023 - 4029, English[Refereed]Scientific journal
- Recombinant yeast strains highly tolerant to formic acid during xylose fermentation were constructed. Microarray analysis of xylose-fermenting Saccharomyces cerevisiae strain overexpressing endogenous xylulokinase in addition to xylose reductase and xylitol dehydrogenase from Pichia stipitis revealed that upregulation of formate dehydrogenase genes (FDH1 and FDH2) was one of the most prominent transcriptional events against excess formic acid. The quantification of formic acid in medium indicated that the innate activity of FDH was too weak to detoxify formic acid. To reinforce the capability for formic acid breakdown, the FDH1 gene was additionally overexpressed in the xylose-metabolizing recombinant yeast. This modification allowed the yeast to rapidly decompose excess formic acid. The yield and final ethanol concentration in the presence of 20 mM formic acid is as essentially same as that of control. The fermentation profile also indicated that the production of xylitol and glycerol, major by-products in xylose fermentation, was not affected by the upregulation of FDH activity.SPRINGER, May 2011, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 90(3) (3), 997 - 1004, English[Refereed]Scientific journal
- Jan. 2011, Microbial Cell Factories, 10(1), 2, EnglishMetabolic pathway engineering based on metabolomics confers acetic and formic acid tolerance to a recombinant xyloseo-fermenting strain ofSaccharomyces cerevisiae[Refereed]Scientific journal
- A yeast with the xylose isomerase (XI) pathway was constructed by the multicopy integration of XI overexpression cassettes into the genome of the Saccharomyces cerevisiae MT8-1 strain. The resulting yeast strain successfully produced ethanol from both xylose as the sole carbon source and a mixed sugar, consisting of xylose and glucose, without any adaptation procedure. Ethanol yields in the fermentation from xylose and mixed sugar were 61.9% and 62.2% of the theoretical carbon recovery, respectively. Knockout of GRE3, a gene encoding nonspecific aldose reductase, of the host yeast strain improved the fermentation profile. Not only specific ethanol production rates but also xylose consumption rates was improved more than twice that of xylose-metabolizing yeast with the XI pathway using GRE3 active yeast as the host strain. In addition, it was demonstrated that xylitol in the medium exhibits a concentration-dependent inhibition effect on the ethanol production from xylose with the yeast harboring the XI-based xylose metabolic pathway. From our findings, the combination of XI-pathway integration and GRE3 knockout could be result in a consolidated xylose assimilation pathway and increased ethanol productivity.SPRINGER, Nov. 2010, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 88(5) (5), 1215 - 1221, English[Refereed]Scientific journal
- Nov. 2010, Applied Microbiology and Biotechnology, 88, 1215-1221, EnglishConstruction of a xylose-metabolizing Saccharomyces cerevisiae by integration of xylose isomerase gene into the genome and investigation of the effect of xylITOl on fermentation[Refereed]Scientific journal
- Sep. 2010, Applied Microbiology and Biotechnology, 88(1), 381-388, EnglishDirect ethanol production from cellulosic materials at high temperature using the thermotolerant yeast Kluyveromyces marxianus displaying cellulolytic enzymes.; Applied Microbiology and Biotechnology[Refereed]Scientific journal
- Aug. 2010, Applied Microbiology and Biotechnology, 87(5), 1975-1982, EnglishCo-fermentation of cellobiose and xylose using beta-glucosidase displaying diploid industrial yeast strain OC-2[Refereed]Scientific journal
- May 2010, Biotechnology Journal, 5, 449-455, EnglishEthanol production from cellulosic materials using cellulase expressing yeast[Refereed]Scientific journal
- Mar. 2010, Journal of Experimental Botany, 61(4), 1041-1051, EnglishMetabolic turnover analysis by a combination of in vivo C-13-lABEling from (CO2)-C-13 and metabolic profiling with CE-MS/MS reveals rate-limiting steps of the C-3 photosynthetic pathwaty in Nicotiana tabacum leaves[Refereed]Scientific journal
- Metabolic pathway engineering by plastid transformation is a powerful tool for production of compounds in higher plantsPlastid transformation is a powerful tool for the production of useful compounds in higher plants through metabolic engineering, because it has many advantages over conventional nuclear transformation: high-level foreign protein accumulation, no need for a transit peptide, absence of gene silencing, and convenient transgene stacking in an operon. Plastid transformation has recently yielded remarkable results in the production of highly valued biopharmaceutical proteins and in conferring herbicide and insect resistance. Metabolic pathway engineering by plastid transformation has also produced higher levels of useful compounds than nuclear transformation. furthermore, recent reports have shown the functional regulation of transgene expression from the plastid genome. In this review, we have focused on the progress of plastid transformation in material production from the aspect of biosynthetic pathway engineering, discussing the issues for future expansion of plastid transformation.JAPANESE SOC PLANT CELL & MOLECULAR BIOL, Mar. 2009, PLANT BIOTECHNOLOGY, 26(1) (1), 39 - 46, English[Refereed]Scientific journal
- Sep. 2008, The Plant Journal, Vol 55. No. 5, pp. 857-868, EnglishBiosynthesis of astaxanthin in tobacco leaves by transplastomic engineering[Refereed]Scientific journal
- May 2008, Journal of Bioscience and Biotechnology, Vol 105. No. 5, pp. 518-526, EnglishOverexpression of 1-deoxy-D-xylulose-5-phosphate reductoisomerase gene in chloroplast contributes to increment of isoprenoid production[Refereed]Scientific journal
- Mar. 2008, Tetrahedron Letters, Vol 49. No. 20, pp. 3294-3296, English4-Ketoantheraxanthin, a novel carotenoid produced by the combination of the bacterial enzyme b-carotene ketolase CrtW and endogeneous carotenoid biosynthetic enzymes in hihger plants[Refereed]Scientific journal
- Aug. 2004, Journal of Biotechnology, Vol 111. No. 3, pp. 241-251, EnglishExpression of fungal pectin methylesterase in transgenic tobacco leads to alteration cell wall metabolism and a dwarf phenotype[Refereed]Scientific journal
- Mar. 2004, Analytical Chemistry, Vol 76. No. 5, pp. 1500-1506, EnglishReal-time quantification of methanol in plants using a hybrid alcohol oxidase-peroxidase biosensor[Refereed]Scientific journal
- Dec. 2003, Journal of Biotechnology, Vol 106. No. 1, pp. 45-52, EnglishMethanol production is enhanced by expression of an Aspergillus niger pectin methylesterase in tobacco cells[Refereed]Scientific journal
- Nov. 2001, Bioorganic & Medicinal Chemistry Letters, Vol. 11, No. 22, pp. 2927-2930, EnglishSELEX for tubulin affords specific T-rich DNA aptamers[Refereed]Scientific journal
- Jan. 2023, JST news, 1, 10 - 11, Japanese特集 CO2から有用物質を生産し低炭素社会実現へ:AIと代謝工学を組み合わせ生産株開発を加速
- Oct. 2022, 日本生物工学会100年史, 58 - 60, Japanese第4章 生物工学のこれから これからの生物育種 -代謝工学による育種のこれから-Introduction scientific journal
- Oct. 2022, 日本生物工学会100年史, 29 - 31, Japanese第3章 生物工学の研究100年 育種技術 -突然変異から代謝工学へ-Introduction scientific journal
- Apr. 2022, 化学工学 [特集]「生物機能を利用したモノづくり」に貢献するプロセス強化, 86(4) (4), 157 - 160, Japanese微生物の高速育種を実現するスマートセル創出プラットフォームIntroduction scientific journal
- Mar. 2022, オレオサイエンス, 22(3) (3), 99 - 105, Japanese進化する酵母細胞表層工学 -バイオインターフェイスの高度利用に向けた技術戦略-Introduction scientific journal
- Sep. 2021, 生物工学会誌, 99(9) (9), 456 - 460, Japanese光合成メタボロミクスの物質生産への応用
- Aug. 2021, 生物工学会誌, 99(8) (8), 403 - 403, Japanese特集 藻類バイオマス利用のための新しい生物工学
- Feb. 2020, 化学工学, 84(2) (2), 51, Japaneseスマートセルインダストリーの形成に向けたバイオ×デジタルの技術開発Introduction scientific journal
- Jan. 2020, バイオサイエンスとインダストリー, 78(1) (1), 32 - 33, Japanese新たに設計した代謝経路を用いた大腸菌によるアルカロイド高生産Introduction scientific journal
- Sep. 2019, バイオサイエンスとインダストリー, 77(5) (5), 410 - 411, Japaneseスマートセル開発に資するメタボロミクス技術Introduction scientific journal
- Sep. 2019, バイオサイエンスとインダストリー, 77(5) (5), 390 - 392, Japanese細胞表層提示酵母を利用したクラフトパルプからのキシリトール生産Introduction scientific journal
- Previous studies have utilized monoamine oxidase (MAO) and L-3,4-dihydroxyphenylalanine decarboxylase (DDC) for microbe-based production of tetrahydropapaveroline (THP), a benzylisoquinoline alkaloid (BIA) precursor to opioid analgesics. In the current study, a phylogenetically distinct Bombyx mori 3,4-dihydroxyphenylacetaldehyde synthase (DHPAAS) is identified to bypass MAO and DDC for direct production of 3,4-dihydroxyphenylacetaldehyde (DHPAA) from L-3,4-dihydroxyphenylalanine (L-DOPA). Structure-based enzyme engineering of DHPAAS results in bifunctional switching between aldehyde synthase and decarboxylase activities. Output of dopamine and DHPAA products is fine-tuned by engineered DHPAAS variants with Phe79Tyr, Tyr80Phe and Asn192His catalytic substitutions. Balance of dopamine and DHPAA products enables improved THP biosynthesis via a symmetrical pathway in Escherichia coli. Rationally engineered insect DHPAAS produces (R,S)-THP in a single enzyme system directly from L-DOPA both in vitro and in vivo, at higher yields than that of the wild-type enzyme. However, DHPAAS-mediated downstream BIA production requires further improvement.01 May 2019, Nature communications, 10(1) (1), 2015 - 2015, English, International magazine
- Mar. 2019, 生物工学会誌, 97(3) (3), 150 - 151, JapaneseBranch SpiritIntroduction scientific journal
- Oct. 2018, 生物工学会, 96(10) (10), 598 - 601, Japaneseチャンスを掴むための準備, バイオ系のキャリアデザインIntroduction scientific journal
- Nov. 2017, アグリバイオ, 1(12) (12), 24 - 29, Japaneseバイオリファイナリー実現を加速する先端バイオ技術と情報技術の融合Introduction scientific journal
- AMER SOC MICROBIOLOGY, Feb. 2017, MICROBIOLOGY RESOURCE ANNOUNCEMENTS, 5(5) (5), EnglishOthers
- 日本農芸化学会 ; 1962-, Feb. 2017, 化学と生物, 55(2) (2), 88 - 97, Japanese多彩な戦略で挑むシアノバクテリア由来の燃料生産 持続可能な第三世代バイオ燃料生産の最前線Introduction scientific journal
- Nov. 2016, レーザー研究, 11, 731 - 734, Japanese微細藻類を利用したバイオ燃料とカロテノイド類の生産Introduction scientific journal
- Nov. 2016, 生物工学, 94(11) (11), 698 - 700, Japanese原子間力顕微鏡による酵母細胞表層セルラーゼの局在評価[Refereed]Introduction scientific journal
- シーエムシー出版, Oct. 2016, バイオマスエネルギーの技術と市場, 第4章, 33 - 45, Japanese次世代バイオエタノール製造技術[Refereed]Introduction scientific journal
- 2016, Journal of Environmental Biotechnology, 16(1) (1), 51 - 58, Japanese代謝プロファイリング法の微生物育種技術への応用Introduction scientific journal
- 日本生物工学会, 2015, 日本生物工学会大会講演要旨集, 67, 250 - 250, English2S-Ca04 Development of microbial cell factories for biorefinery
- 2015, 日本生物工学会大会講演要旨集, 67th好気・低濃度グルコース条件下におけるSaccharomyces cerevisiaeの転写解析
- 2015, 日本生物工学会大会講演要旨集, 67thSaccharomyces cerevisiaeの糖代謝における転写制御ネットワークの予測
- 日本生物工学会, 2014, 日本生物工学会大会講演要旨集, 66, 206 - 206, Japanese3P-045 Effects of hydrolysate of lignocellulosic biomass on phenyllactate fermentation by recombinant Escherichia coli
- 日本生物工学会, 2014, 日本生物工学会大会講演要旨集, 66, 205 - 205, Japanese3P-044 Improvement of CHO cells useful in bio-pharmaceutical development, construction of data leading to the culture state elucidation
- 日本生物工学会, 2014, 日本生物工学会大会講演要旨集, 66, 51 - 51, Japanese1P-134 Development of the novel secretion signal sequence for highly-efficient cell surface display and secretory production of proteins by yeasts
- 日本生物工学会, 2014, 日本生物工学会大会講演要旨集, 66, 246 - 246, Japanese3P-205 Metabolite profiling of oceanic cyanobacteria under salt stress condition
- 日本生物工学会, 2014, 日本生物工学会大会講演要旨集, 66, 252 - 252, Japanese3P-229 Effect of PHO13 gene disruption on xylose production in xylose-fermenting yeast harboring xylose isomerase gene
- 日本生物工学会, 2014, 日本生物工学会大会講演要旨集, 66, 139 - 139, Japanese2P-132 Development of lipid-producing system depending on estimation of cell composition and metabolic analysis for marine green alga Chlamydomonas sp. JSC4
- 日本生物工学会, 2014, 日本生物工学会大会講演要旨集, 66, 140 - 140, English2P-136 Effect of illumination coupled with nitrogen depletion on biodiesel production of a marine microalga Chlamydomonas sp. JSC4 :
- 日本生物工学会, 2014, 日本生物工学会大会講演要旨集, 66, 130 - 130, Japanese2P-095 High-level ethanol production from cyanobacterium Spirulina by amylase displayed yeast
- 日本生物工学会, 2014, 日本生物工学会大会講演要旨集, 66, 98 - 98, Japanese2A-Dp01 Development of microbe breeding technology based on metabolic profiling
- 日本生物工学会, 2014, 日本生物工学会大会講演要旨集, 66, 44 - 44, Japanese1P-110 Detoxification of furfural in Corynebacterium glutamicum under aerobic and anaerobic conditions
- 日本生物工学会, 2014, 日本生物工学会大会講演要旨集, 66, 177 - 177, Japanese3S-Ba05 Challenge for liquid fuel production from algae based on metabolic analysis
- 2014, 日本生物工学会大会講演要旨集, 66thキシロース資化性酵母における遺伝子発現プロファイルの経時変化解析
- 2014, 日本生物工学会大会講演要旨集, 66thKluyveromyces marxianusの好気条件での糖代謝系の解析
- Aug. 2013, 安全工学, 52(4) (4), 249 - 255, Japaneseバイオリファイナリー社会に向けた燃料・化学品生産Introduction scientific journal
- 日本生物工学会, Jun. 2013, 生物工学会誌, 91(6) (6), 314 - 318, Japanese革新的なものづくり実現のための「合成生物工学」Introduction scientific journal
- 日本生物工学会, 2013, 日本生物工学会大会講演要旨集, 65, 43 - 43, Japanese1P-104 Identification of genes related to improved ethanol production from xylose in presence of acetate and formate by regulation of metabolism in Saccharomyces cerevisiae
- 日本生物工学会, 2013, 日本生物工学会大会講演要旨集, 65, 64 - 64, Japanese1P-185 Development of high lipid producing system by green alga Chlamydomonas orbicularis under sea salt condition
- 日本生物工学会, 2013, 日本生物工学会大会講演要旨集, 65, 66 - 66, English1P-196 Phototrophic cultivation of a marine microalga Chlamydomonas orbicularis for CO_2 fixation and biodiesel production: Effect of medium composition, nitrogen depletion, and sea salt concentration :
- 日本生物工学会, 2013, 日本生物工学会大会講演要旨集, 65, 64 - 64, Japanese1P-186 Glycogen production for biorefinery using oceanic cyanobacteria
- 日本生物工学会, 2013, 日本生物工学会大会講演要旨集, 65, 54 - 54, Japanese1P-146 Improvement of ethanol production from xylose by regulation of the PHO pathway in Saccharomyces cerevisiae
- 日本生物工学会, 2013, 日本生物工学会大会講演要旨集, 65, 65 - 65, Japanese1P-191 The effect of sea-salts on the cell-growth and photosynthesis in a remarkable carbon feedstock Spirulina
- 日本生物工学会, 2013, 日本生物工学会大会講演要旨集, 65, 63 - 63, Japanese1P-184 Development of novel yeast cell surface display system for consolidated bioethanol production from lignocellulosic biomass
- 日本生物工学会, 2013, 日本生物工学会大会講演要旨集, 65, 141 - 141, Japanese2P-148 Sugar concentration in liquid hydrolysate of dilute acid pretreated rice straw by nanofiltration
- 2013, 日本生物工学会大会講演要旨集, 65th統合バイオプロセスによる環境調和型セルロース系エタノール生産に資する前処理技術の開発
- 2013, 日本農芸化学会大会講演要旨集(Web), 2013発酵阻害物存在下におけるキシロースからのエタノール発酵向上酵母の構築
- Mar. 2012, 第53回日本植物生理学会, JapaneseCyanobacterium Synechocystis sp. PCC6803 におけるO2存性オルタナティブ・エレクトロン・フロー (AEF)の生理的役割
- 日本生物工学会, 2012, 日本生物工学会大会講演要旨集, 64, 65 - 65, Japanese2Gp16 Metabolome analysis of filamentous fungus Rhizopus oryzae showing different behaviors of lipase secretion
- 日本生物工学会, Oct. 2011, 生物工学会誌, 89(10) (10), 585 - 588, Japanese有用希少イノシトールのバイオアベイラビリティーとバイオコンバージョン生産Introduction scientific journal
- Oct. 2011, ブレインテクノニュース, 144, 17 - 22, JapaneseCBP用スーパー酵母を用いたバイオエタノール生産技術の開発Introduction scientific journal
- 化学工業社, Sep. 2011, ケミカルエンジニヤリング, 56(9) (9), 661 - 668, Japaneseバイオマスからの効率的なバイオ燃料の生産技術 (特集 本格化するバイオマスエネルギー開発)
- Sep. 2011, ケミカルエンジニヤリング, 56(9) (9), 5 - 12, Japaneseバイオマスからの効率的なバイオ燃料の生産技術Introduction scientific journal
- 一般社団法人日本土壌肥料学会, 08 Aug. 2011, 日本土壌肥料学会講演要旨集, (57) (57), 329 - 329, Japanese11 シアノバクテリアでのオルタナティブ・エレクトロン・フロー(AEF)評価系の確立(関西支部講演会,2010年度各支部会講演要旨)
- Apr. 2011, 生物工学会誌, 89(4) (4), 181 - 183, Japanese微細藻類によるバイオリファイナリーIntroduction scientific journal
- 日本生物工学会, 2011, 日本生物工学会大会講演要旨集, 63, 180 - 180, Japanese2Ip14 High efficient ethanol production from hydrothermally-processed rice straw using cellulase-displaying yeast with a rotary fermenter
- 日本生物工学会, 2011, 日本生物工学会大会講演要旨集, 63, 187 - 187, Japanese2Ka04 Reduction of furan derivatives by NADH-dependent Adh1 improves ethanol fermentation using xylose as sole carbon source with Saccharomyces cerevisiae harboring XR-XDH pathway
- Nov. 2010, バイオインダストリー, 11, 6-12, Japaneseバイオマスからのバイオ燃料生産に向けた戦略Introduction scientific journal
- Jul. 2010, 生物工学, 88(7), 333-335, Japanese次世代燃料・化成品原料製造に向けたバイオリファイナリー戦略Introduction scientific journal
- 日本ビタミン学会, 25 Apr. 2010, ビタミン, 84(4) (4), 206 - 206, Japanese2-IV-14 D-アラビノースは酵母エリスロアスコルビン酸の内在性前駆体ではない(一般演題,日本ビタミン学会第62回大会発表要旨)
- 日本生物工学会, 2010, 日本生物工学会大会講演要旨集, 22, 87 - 87, Japanese3P-1119 Efficient ethanol production from xylose in the presence of fermentation inhibitors
- 日本生物工学会, 2010, 日本生物工学会大会講演要旨集, 22, 202 - 202, Japanese3S-Ba04 Development of bio-fuel production system using microalgae
- 日本生物工学会, 2010, 日本生物工学会大会講演要旨集, 22, 119 - 119, Japanese2P-2051 Efficient fermentation of xylose to ethanol in the presence of furan compound by recombinant Saccharomyces cerevisiae expressing Alcohol dehydrogenase
- 日本生物工学会, 2010, 日本生物工学会大会講演要旨集, 22, 60 - 60, Japanese3P-1011 Construction of the recombinant xylose fermentation yeast with high β-xylosidase activity from OC-2 triple marker strain as a platform
- The dependency on depleting natural resources is a challenge for energy security that can be potentially answered by bioenergy. Bioenergy is derived from starchy and lignocellulosic biomass in the form of bioethanol or from vegetable oils in the form of biodiesel fuel. The acid and enzymatic methods have been developed for the hydrolysis of biomass and for transesterifiaction of plant oils. However, acid hydrolysis results in the production of unnatural compounds which has adverse effects on yeast fermentation. Recent advancements in the yeast cell surface engineering developed strategies to genetically immobilize amylolytic, cellulolytic and xylanolytic enzymes on yeast cell surface for the production of fuel ethanol from biomass. This review gives an insight in to the recent technological developments in the production of bioenergy, i.e, bioethanol using surface engineered yeast. © 2010 Bentham Science Publishers Ltd.2010, Recent Patents on Biotechnology, 4(3) (3), 226 - 234, English[Refereed]Introduction scientific journal
- The Society for Biotechnology, Japan, Nov. 2009, Journal of bioscience and bioengineering, 108(1) (1), S167, EnglishSB-O5 Dynamic metabolic profiling in plant leaves using in vivo stable isotope labeling(Section XI Systems Biotechnology/Metabolic Engineering)
- SOC BIOSCIENCE BIOENGINEERING JAPAN, Nov. 2009, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 108, S167 - S167, EnglishSummary international conference
- The Society for Biotechnology, Japan, Nov. 2009, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 108(1) (1), S53, English[Refereed]Summary international conference
- Nov. 2009, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 108, S53, English[Refereed]Summary international conference
- Mar. 2009, ケミカルエンジニヤリング, Vol 54. No. 3, pp. 24-30, Japaneseバイオマスからの化学品生産を目指したバイオプロセスの開発Introduction scientific journal
- 日本生物工学会, 2009, 日本生物工学会大会講演要旨集, 21, 292 - 292, Japanese3S15a04 Bioproduction through synthetic bioengineering
- 2008, 生化学13CO2を用いた同位体標識による葉内代謝物質のターンオーバー解析
- Dynamic metabolic profiling using in vivo stable isotope labeling生体試料に含まれる代謝産物の量的情報をMSやNMRを利用して網羅的に解析する代謝プロファイリング技術は,微量な中間代謝物質を含む多様な化合物の一斉分析を可能にしたが,ここで得られる情報は代謝物質を抽出した時点での蓄積量のスナップショットであった.生体内で代謝物質が動的定常状態にある時,総量は変わらずに一定の同じ速度で分解と合成が行われて入れ替わっており,新規合成される物質の存在比率を知るためには安定同位体を用いたin vivo標識が必須である.代謝プロファイリングと安定同位体標識法の組合せを利用した「動的代謝プロファイリング」は,代謝産物の量的変動を網羅的に観測することを可能にする.植物の環境変化やストレスに対する応答はシステマティックな代謝変動に基づくことが予想され,「動的代謝プロファイリング」はこうした複雑な応答機構の解明に寄与することが期待できる.Metabolic profiling technique has enabled to analyze the level of various metabolites including minor intermediates by using MS or NMR simultaneously, but the information available from the technique was snapshot in vivo at the time of sampling metabolites. In the dynamic steady state, metabolites are replaced with newly-synthesized compounds at a constant rate and the total amount of metabolites remains unchanged. Therefore, in vivo labeling with stable isotopes is necessary for the determination of turnover rates of metabolites. Dynamic metabolic profiling formed by the combination of metabolic profiling and stable isotope labeling technique enables to determine changes in metabolic flux exhaustively. In plants, responses to stress or environmental changes would be based on systematic variations in metabolism. The dynamic metabolic profiling would contribute to elucidate the complicated response mechanism.光合成研究法. 北海道大学低温科学研究所, 日本光合成研究会共編北海道大学低温科学研究所 = Institute of Low Temperature Science, Hokkaido University, 2008, Low temperature science, 67, 169 - 174, Japanese
- 公益社団法人日本生物工学会, 25 Aug. 2003, 日本生物工学会大会講演要旨集, 15, JapaneseExpression of Aspergillus niger pectin methylesterase in transgenic tobacco (Nicotiana tabacum) leads to alterations on the level of methanol and a dwarf phenotype
- 公益社団法人日本生物工学会, 10 Jul. 2000, 日本生物工学会大会講演要旨集, 12, Japanese
- Joint work, 第9章 バイオマス燃料生産の技術動向, シーエムシー出版, Feb. 2022クリーンエネルギーの技術と市場 2022
- 第2章 醸造の科学, 最近の育種技術, 朝倉書店, Jun. 2021醸造の辞典
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- Metabolic Engineering for Carotenoid Production Using Eukaryotic Microalgae and Prokaryotic Cyanobacteria, Springer, 2021Carotenoids: Biosynthetic and Biofunctional Approaches
- Joint work, 第Ⅲ編 生物プロセス, 第2章 ラン藻によるバイオコハク酸生産, シーエムシー, Jul. 2020脱石油に向けたCO2資源化技術―化学・生物プロセスを中心に―
- Joint work, 第3章 7. 組織内の分子を網羅的に測る技術, シーエムシー・リサーチ, Jun. 2019骨格筋研究を核とした筋スマート社会
- Joint work, シーエムシー出版, Jun. 2018, Japanese第9章 油脂酵母による油脂発酵生産性改善へ向けた技術開発, スマートセルインダストリー -微生物細胞を用いた物質生産の展望-, 第3編 産業応用へのアプローチScholarly book
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- Joint work, シーエムシー出版, Jun. 2018, Japaneseスマートセルインダストリー –微生物細胞を用いた物質生産の展望–(第1編 ハイスループット合成・分析・評価技術,第2章 ハイスループット微生物構築・評価技術,第1節 「微生物を用いた物質生産とハイスループット微生物構築技術」)Scholarly book
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- Joint work, Wiley-VCH, Feb. 2018, EnglishEmerging Areas in Bioengineering:Whole cell biocatalysts using enzymes displayed on yeast cell surfaceScholarly book
- Joint work, Wley, Nov. 2016, EnglishIndustrial Biotechnoligy : Products and Processes, Chapter 5, Production of fuels and chemicals from biomass by integrated bioprocessesScholarly book
- Joint work, 文永堂出版, Aug. 2016, Japanese応用微生物学,第10章 低炭素化社会への取組み,バイオ燃料Scholarly book
- Joint work, Wiley, Apr. 2014, EnglishBioprocessing of Renewable Resources to Commodity Bioproducts, Chapter 8, Ethanol production from yeastsScholarly book
- Joint work, 丸善出版, Jan. 2014, Japanese化学便覧 応用化学編Scholarly book
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- Joint work, シーエムシー出版, May 2013, Japaneseリサイクルバイオテクノロジーの最前線, 第I編 第1章, 1 未来型CBP法によるバイオエタノール生産技術の展開Scholarly book
- Joint work, 化学同人, Oct. 2012, Japaneseひらく、ひらく「バイオの世界」Scholarly book
- Joint work, シーエムシー出版, Jul. 2012, Japanese微細藻類によるエネルギー生産と事業展望Scholarly book
- Joint work, エヌ・ティー・エス, Jul. 2012, Japanese藻類ハンドブックScholarly book
- Joint work, シーエムシー出版, Apr. 2012, Japanese合成生物工学の隆起Scholarly book
- Joint work, 三共出版, Mar. 2012, Japanese微生物機能学Scholarly book
- Joint work, シーエムシー出版, Mar. 2012, Japaneseバイオマス分解酵素研究の最前線, 第7編, 第28章 セルラーゼ細胞表層提示酵母を用いたバイオマス変換Scholarly book
- Joint work, シーエムシー出版, Mar. 2012, Japaneseバイオマス分解酵素研究の最前線, 序章, 2 バイオマス分解酵素研究の新たな展開Scholarly book
- Joint work, Springer, 2012, EnglishSystem Metabolic EngineeringScholarly book
- Joint work, シーエムシー出版, Dec. 2010, JapaneseエコバイオリファイナリーScholarly book
- Joint work, シーエムシー出版, Apr. 2010, Japaneseグリーンバイオケミストリーの最前線Scholarly book
- Joint work, Humana Press, Feb. 2010, EnglishMethods in Molecular BiologyScholarly book
- Joint work, フロンティア出版, Jan. 2010, Japanese次世代バイオエタノール生産の技術革新と事業展開, 第7章, 4 アーミング酵母を用いた統合プロセスによるバイオエタノール生産Scholarly book
- Joint work, フロンティア出版, Jan. 2010, Japanese次世代バイオエタノール生産の技術革新と事業展開, 第10章, 7 アーミング技術を応用した新規セルロース分解システムの構築Scholarly book
- Joint work, シーエムシー出版, Apr. 2009, Japanese第二世代バイオ燃料の開発と応用展開Scholarly book
- Joint work, 生物研究社, Mar. 2009, Japanese水環境の今と未来 藻類と植物のできることGeneral book
- Joint work, 北海道大学低温科学研究所・日本光合成研究会共編, Mar. 2009, Japanese光合成研究法Scholarly book
- レーザー学会学術講演会第41回年次大会シンポジウム, Jan. 2021, Japaneseポストコロナ時代の産学連携について考える
- 第一回先端膜工学研究推進機構特定テーマフォーラム~医薬・バイオプロセスにおける膜利用の現状と将来展望~, Dec. 2020, Japaneseバイオプロセスにおける膜利用の現状と将来展望
- 2nd ASBA Webinar, Dec. 2020, EnglishMetabolomics-based engineering biology for microbial bio-production with sustainability
- CBI学会2020年大会, Oct. 2020, Japaneseハイスループットプラットフォームの開発による微生物スマートセルの構築
- JPC関西特別報告会, Oct. 2020, Japanese微細藻類を利用した物質生産プロセスへのLEDの応用
- Laboratory Automation勉強会, Aug. 2020, Japaneseメタボローム解析の自動化に向けた挑戦
- 質量分析インフォマティクス研究会 第5回ワークショップ, Aug. 2020, Japaneseバイオ×デジタルの技術融合による有用微生物「スマートセル」開発への挑戦[Invited]
- 日本農芸化学会2020年度大会, Japaneseスマートセルインダストリーに資するハイスループット微生物構築・評価技術の開発
- NEDOフェスタin関西 2019, Dec. 2019, Japanese, Domestic conferenceスマートセル開発に資するハイスループット微生物構築・評価技術の概要[Invited]Public discourse
- 日本農芸化学会関西支部 支部例会(第511回講演会), Dec. 2019, Japanese, Domestic conference酵母Saccharomyces cerevisiaeの鉄代謝改変に基づくキシロースからの1,2,4-ブタントリオール発酵生産Oral presentation
- 日本農芸化学会関西支部 支部例会(第511回講演会), Dec. 2019, Japanese, Domestic conferenceスマートセルインダストリーに資するメタボローム解析技術の開発と応用[Invited]Invited oral presentation
- 藍藻の分子生物学2019, Nov. 2019, Japanese, Domestic conference藍藻の動的メタボローム解析技術の開発と応用[Invited]Invited oral presentation
- Bio Japan 2019, Oct. 2019, Japanese, Domestic conferenceバイオ×デジタルによるスマートセル開発への挑戦[Invited]Nominated symposium
- Metabolic Engineering Summit 2019, Oct. 2019, English, International conferenceMetabolomics-based engineering for smart cell developmentOral presentation
- 2019 Asian Synthetic Biology Association (ASBA) Meeting, Oct. 2019, English, International conferenceMetabolomics-based engineering biology for bio-productionOral presentation
- 第61回天然有機化合物討論会, Sep. 2019, English, Domestic conferenceRe-design, synthesis, and bio-analysis of sulfosialic acids as covalent inhibitors of neuraminidasePoster presentation
- 2019年度NEDOスマートセルプロジェクト技術セミナー, Sep. 2019, Japanese, Domestic conferenceスマートセル開発に資するハイスループット微生物構築・評価技術の概要Invited oral presentation
- 第71回日本生物工学会大会, Sep. 2019, Japanese, 岡山大学, Domestic conference窒素源存在下で油脂を高蓄積する海洋性クラミドモナス変異株の選択的育種Oral presentation
- 第71回日本生物工学会大会, Sep. 2019, Japanese, 岡山大学, Domestic conference食農産業における LED の利活用技術の開発Oral presentation
- 第71回日本生物工学会大会, Sep. 2019, Japanese, 岡山大学, Domestic conference細胞表層提示システムに適した宿主酵母の開発Oral presentation
- 第71回日本生物工学会大会, Sep. 2019, Japanese, 岡山大学, Domestic conference海洋性クラミドモナス油脂高蓄積変異株における油脂・カロテノイド増産メカニズムの解明Oral presentation
- 第71回日本生物工学会大会, Sep. 2019, Japanese, 岡山大学 津島キャンパス, Domestic conferencePichia pastorisにおいて分泌シグナル配列の1アミノ酸置換はタンパク質分泌生産量を劇的に増大させるOral presentation
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- 第70回日本生物工学会大会, Sep. 2018, Japanese, 日本生物工学会, 関西大学 千里山キャンパス, Domestic conference鉄硫黄タンパク質高活性化に基づくキシロースからの1,2,4-ブタントリオール生産技術の開発Poster presentation
- 第70回日本生物工学会大会, Sep. 2018, Japanese, 関西大学 千里山キャンパス, Domestic conference組換え出芽酵母Saccharomyces cerevisiaeによるキシロースからのグルタチオン生産Poster presentation
- 第70回日本生物工学会大会, Sep. 2018, Japanese, 日本生物工学会, 関西大学 千里山キャンパス, Domestic conference接合型変換を利用したセルラーゼ群表層提示酵母の糖化活性及び発酵阻害物耐性向上Poster presentation
- 第70回日本生物工学会大会, Sep. 2018, Japanese, 日本生物工学会, 関西大学 千里山キャンパス, Domestic conference酵母Scheffersomyces stipitisを用いたレスベラトロール生産プロセスの開発Poster presentation
- 第70回日本生物工学会大会, Sep. 2018, Japanese, 日本生物工学会, 関西大学 千里山キャンパス, Domestic conference酵母Scheffersomyces stipitisを宿主とした新規細胞表層提示システムの開発Poster presentation
- 第70回日本生物工学会大会, Sep. 2018, Japanese, 日本生物工学会, 関西大学 千里山キャンパス, Domestic conference遺伝子組換え効率向上に向けたDNAリガーゼIV欠損Pichia pastoris株の開発Oral presentation
- 第70回日本生物工学会大会, Sep. 2018, Japanese, 関西大学 千里山キャンパス, Domestic conferenceラン藻窒素代謝メカニズムの解明と物質生産への応用Poster presentation
- 第70回日本生物工学会大会, Sep. 2018, Japanese, 関西大学 千里山キャンパス, Domestic conferenceスマートセル創出プラットフォームに資するメタボローム解析技術の開発Oral presentation
- 第70回日本生物工学会大会, Sep. 2018, Japanese, 日本生物工学会, 関西大学 千里山キャンパス, Domestic conferencePichia pastorisにおけるセントロメアDNA配列を用いた自律複製型プラスミドベクターの開発Oral presentation
- The International Symposium on Phototrophic Prokaryotes (ISPP), Aug. 2018, English, the Life Sciences Centre, The University of Buritish Columbia, International conferenceTemperature dependent succinate biosynthesis concurrent with alteration in central metabolism turnover in synechocystis SP.PCC6803Oral presentation
- 第13回 アジレントメタボロミクスセミナー, Jul. 2018, Japanese, 慶応義塾大学 三田キャンパス, Domestic conference動的メタボロミクスの開発とスマートセルインダストリーへの展開[Invited]Invited oral presentation
- 第20回 新産業技術促進検討会, Jul. 2018, Japanese, TKPガーデンシティPREMIUM神保町 プレミアムボールルーム, Domestic conferenceスマートセル創出プラットフォームに資するハイスループット微生物構築・評価技術の開発Oral presentation
- Metabolic Engineering 12, Jun. 2018, English, The Westin Grand Munich, International conferenceDynamic Metabolomics of Crabtree-Negative Yeast Kluyveromyces MarxianusOral presentation
- The 8th International Conference on Algal Biomass, Biofuels and Bioproducts (AlgalBBB 2018), Jun. 2018, English, Motif Seattle, International conferenceConverting Chlamydomonas sp. JSC4 lipids to biodiesel via biocatalysis with Fusarium heterosporum lipase-expressing Aspergillus oryzae whole-cellPoster presentation
- ライフサイエンス技術部会・反応分科会講演会講演会「スマートセルインダストリーに関する研究開発動向」, Feb. 2018, Japanese, Domestic conferenceスマートセルインダストリーの創出に資する微生物育種プラットフォームの開発[Invited]Public discourse
- The 9th International Symposium of Innovative BioProduction Kobe (iBioK), Feb. 2018, English, Kobe, Japan, International conferenceEnhanced protein secretion by accumulation of novel effective factors in Pichia pastorisPoster presentation
- The 9th International Symposium of Innovative BioProduction Kobe (iBioK), Feb. 2018, English, Kobe, Japan, International conferenceA new autonomous replicating plasmid vector containing Pichia pastoris centromeric DNAPoster presentation
- シンポジウム「微細藻類を利用したバイオ燃料生産の研究開発動向」, Jan. 2018, Japanese, オフィス東京, Domestic conference藻類オイル生産の実用化に向けた代謝メカニズム解析の重要性Oral presentation
- 5th Asian Conference on Biomass Science, Jan. 2018, English, 日本エネルギー学会, 仙台市, International conferenceEfficient Xylitol Production from Kraft Pulp by a Cell Surface Engineered Strain ofSaccharomyces cerevisiaePoster presentation
- 静岡大学グリーン科学研究所 第4回シンポジウム, Nov. 2017, Japanese, Domestic conferenceEngineering Biologyによるバイオリファイナリーの構築とスマートセルインダストリーへの展開[Invited]Invited oral presentation
- 第11回メタボロームシンポジウム, Nov. 2017, Japanese, ホテル阪急エキスポパーク, Domestic conference動的メタボロミクスの開発とスマートセルインダストリーへの展開Oral presentation
- The 8th Kobe University Brussels European Centre Symposium, Nov. 2017, English, Vrije Universiteit Brussel U-Residence, International conferenceDevelopment of dynamic metabolomics and its application to metabolic engineeringOral presentation
- BioJapan 2017, Oct. 2017, Japanese, パシフィコ横浜, Domestic conferenceスマートセルを創出する合成バイオプラットフォームの開発Oral presentation
- 第69回日本生物工学会大会, Sep. 2017, Japanese, 日本生物工学会, 東京都新宿区, Domestic conference油脂高蓄積クラミドモナス株の選抜育種によるバイオ燃料生産「明暗周期問題」の克服Oral presentation
- 第31回カロテノイド研究談話会, Sep. 2017, Japanese, 京都薬科大学, Domestic conference代謝工学的手法による海洋性ラン藻でのアスタキサンチン生産Poster presentation
- 第69回日本生物工学会大会, Sep. 2017, Japanese, 日本生物工学会, 東京都新宿区, Domestic conference代謝工学による黄色天然色素ルテイン高蓄積クラミドモナス株の創出Poster presentation
- 第69回日本生物工学会大会, Sep. 2017, Japanese, 日本生物工学会, 東京都新宿区, Domestic conferenceアスタキサンチン生産能を付与した高増殖性ラン藻のトランスクリプトミクスおよび動的メタボロ ミクスPoster presentation
- 第69回日本生物工学会大会, Sep. 2017, Japanese, 日本生物工学会, 早稲田大学 西早稲田キャンパス, Domestic conferencePichia pastorisのタンパク質分泌生産における新規有用因子の獲得とその蓄積による効果の検証Poster presentation
- 第69回日本生物工学会大会, Sep. 2017, Japanese, 日本生物工学会, 早稲田大学 西早稲田キャンパス, Domestic conferencePichia pastorisにおけるセントロメア配列を含む自律複製型プラスミドの開発Poster presentation
- 第69回日本生物工学会大会, Sep. 2017, Japanese, 日本生物工学会, 東京都新宿区, Domestic conferenceN-アセチルグルコサミン資化性酵母Scheffersomyces stipitisの基質別代謝特性の解析Oral presentation
- 第69回日本生物工学会大会, Sep. 2017, Japanese, 日本生物工学会, 東京都新宿区, Domestic conferenceEnhanced cell surface engineering of Saccharomyces cerevisiae and optimization of the fermentation process for efficient xylitol production from kraft pulpOral presentation
- 第69回日本生物工学会大会 国際シンポジウム, Sep. 2017, English, 早稲田大学 西早稲田キャンパス, International conferenceChallenge to development of “smart” microorganisms based on information analysis and its application to production of highly functional compoundsOral presentation
- 合成生物工学シンポジウム, Aug. 2017, Japanese, 神戸大学百年記念館六甲ホール, Domestic conferenceスマートセルを創出する合成バイオプラットフォームの開発と応用への挑戦Oral presentation
- The 18th International Symposium on Carotenoids 2017, Jul. 2017, English, The Lucerne Hall, International conferenceEnhancement in central metabolism and carotenoid biosynthesis by photosynthetic astaxanthin production in the recombinant marine cyanobacteriumOral presentation
- SB7.0 The Seventh International Meeting on Synethetic Biology, Jun. 2017, English, University Cultural Center, International conferenceCell surface engineering of Saccharomyces cerevisiae for biomass breakdownOral presentation
- エネルギー・資源技術部会 バイオマス分科会 講演会「バイオマス資源変換触媒の研究動向」, Apr. 2017, Japanese, Domestic conference進化した細胞表層工学によるバイオマス変換プロセスの開発と機能性物質生産への新展開[Invited]Invited oral presentation
- 日本農芸化学2017年度大会, Mar. 2017, Japanese, 日本農芸化学会, 京都市, Domestic conference変異チオール酸化酵素による酵母の酸化型グルタチオン 生産Oral presentation
- 日本農芸化学2017年度大会, Mar. 2017, Japanese, 日本農芸化学会, 京都市, Domestic conference代謝工学によるアスタキサンチン産生海洋性ラン藻の 開発と代謝メカニズムの解明Oral presentation
- 日本農芸化学会2017年度大会, Mar. 2017, Japanese, 京都女子大学, Domestic conferenceバイオ燃料生産に向けた耐塩性緑藻の進化工学的育種と 塩ストレス耐性による油脂生産低下メカニズムの解明Oral presentation
- 日本農芸化学2017年度大会, Mar. 2017, Japanese, 日本農芸化学会, 京都市, Domestic conferenceバイオ燃料生産に向けた耐塩性緑藻の進化工学的育種と 塩ストレス耐性による油脂生産低下メカニズムの解明Oral presentation
- 日本農芸化学2017年度大会, Mar. 2017, Japanese, 日本農芸化学会, 京都女子大学, Domestic conferenceゲノム編集による Kluyveromyces marxianus の自在な 遺伝子改変Oral presentation
- 日本農芸化学2017年度大会, Mar. 2017, Japanese, 日本農芸化学会, 京都市, Domestic conferenceSaccharomyces cerevisiae による1,2,4-ブタントリオー ル生産技術の開発Oral presentation
- 第12回バイオマス科学会議, Jan. 2017, Japanese, 日本エネルギー学会, 東京都文京区, Domestic conference統合型バイオエタノール生産プロセスの構築に資する改良型酵 母細胞表層提示用遺伝子カセットの開発Oral presentation
- 第21回関西大学先端科学技術シンポジウム, Jan. 2017, Japanese, 関西大学 千里山キャンパス, Domestic conferenceバイオリファイナリーの構築に資する微生物細胞工場の創製[Invited]Invited oral presentation
- 「植物等の生物を用いた高機能品生産技術の開発」キックオフシンポジウム, Nov. 2016, Japanese, ベイサイドホテル アジュール竹芝, Domestic conference日本独自の超高速微生物育種プラットフォーム「スマートセル・ファウンドリ」の開発Oral presentation
- The 9th Asia-Pacific Conference on Algal Biotechnology (APCAB), Nov. 2016, English, Century Park Hotel, International conferenceDevelopment of dynamic metabolic profiling and its application to fuel and chemical production in algaeOral presentation
- 第68回日本生物工学会大会, Sep. 2016, Japanese, 日本生物工学会, 富山市, Domestic conference藍藻 Synechocystis sp. PCC 6803 hox遺伝子変異株のコントロールされた培養条件における有機酸 生産Oral presentation
- 第68回日本生物工学会大会, Sep. 2016, Japanese, 日本生物工学会, 富山市, Domestic conference明暗周期下で効率的に油脂を生産するための海洋性緑藻培養技術の開発Oral presentation
- 第68回日本生物工学会大会, Sep. 2016, Japanese, 日本生物工学会, 富山市, Domestic conference酵母表層への酵素立体的配置技術の開発Oral presentation
- 第68回日本生物工学会大会, Sep. 2016, Japanese, 日本生物工学会, 富山市, Domestic conferenceラン藻スピルリナからの高生産エタノール変換プロセスの開発Oral presentation
- 第68回日本生物工学会大会, Sep. 2016, Japanese, 日本生物工学会, 富山市, Domestic conferenceバイオ医薬生産に向けたCHO細胞培養情報データ解析システムの構築Oral presentation
- 第118回触媒討論会, Sep. 2016, Japanese, 岩手大学, Domestic conferenceバイオリファイナリーの構築に資する微生物細胞工場の創生[Invited]Invited oral presentation
- 第68回日本生物工学会大会, Sep. 2016, Japanese, 日本生物工学会, 富山市, Domestic conferenceN-アセチルグルコサミン資化性酵母Scheffersomyces stipitisの特性解析Oral presentation
- The 14th International Congress on Yeasts (ICY2016), Sep. 2016, English, ICY14組織委員会, 淡路市, International conferenceEnhanced protein secretion by accumulation of novel effective factors in Pichia pastorisPoster presentation
- The 4th Asia-Oceania Algae Innovation Summit (AOAIS2016), Sep. 2016, English, East Lake International Conference Center, Wuhan, China, International conferenceBio-based fuel and chemicalproduction based on dynamic metabolic profiling of cynobacteria and microalgaeOral presentation
- The 14th International Congress on Yeasts (ICY2016), Sep. 2016, English, Awaji, Japan, International conferenceA new autonomous replicating plasmid vector containing centromere DNA sequence of Pichia pastorisPoster presentation
- 生物工学若手研究者の集い夏のセミナー2016, Jul. 2016, Japanese, 生物工学会, 府中市, Domestic conference海洋性ラン藻Synechococcus sp. PCC 7002を用いたアスタキサンチン生産技術の開発Poster presentation
- KMB2016 Annual meeting and international symposium, Jun. 2016, English, International conferenceDevelopment of cell factories of microalgae and cyanobacteria based on metabolic analysis[Invited]Invited oral presentation
- 環境バイオテクノロジー学会2016年度大会, Jun. 2016, Japanese, サテライトキャンパスひろしま, Domestic conference代謝プロファイリング法の紹介と微生物育種技術への応用Oral presentation
- Metabolic Engineering 11, Jun. 2016, English, Metabolic Engineering 11, 淡路市, International conferenceSynthetic Promoter Systems for Controlling Saccharomyces Cerevisiae Gene ExpressionOral presentation
- Metabolic Engineering 11, Jun. 2016, English, Metabolic Engineering 11, 淡路市, International conferenceEngineering of a Novel Cellulose-Adherent Cellulolytic Saccharomyces Cerevisiae for Cellulosic Biofuel ProductionPoster presentation
- Metabolic Engineering 11, Jun. 2016, English, Metabolic Engineering 11, 淡路市, International conferenceDisruption of PHO13 Improves Ethanol Production Via the Xylose Isomerase PathwayOral presentation
- Metabolic Engineering 11, Jun. 2016, English, Metabolic Engineering 11, 淡路市, International conferenceDirect and High-Productive Conversion from a Cyanobacterium Arthrospira Platensis to EthanolOral presentation
- Metabolic Engineering 11, Jun. 2016, English, Metabolic Engineering 11, 淡路市, International conferenceAlternative Inverse Metabolic Engineering Based on Transient Acclimation of Yeast Improves Acid-Containing Xylose Fermentation and Tolerance to Formic and Acetic AcidsOral presentation
- 第340回細胞工学研究会後援会/第220回遺伝子機能解析部門セミナー, Mar. 2016, Japanese, Domestic conference動的代謝プロファイリング技術の開発とバイオリファイナリーへの展開[Invited]Invited oral presentation
- 化学工学会第81年会, Mar. 2016, Japanese, Domestic conferenceAFM力計測を用いた細胞表層提示酵素の特異的検出とマッピング評価Oral presentation
- 日本化学会第96春季年回, Mar. 2016, Japanese, Domestic conferenceAFM力学計測法を利用した細胞表層空間に提示した酵素検出Oral presentation
- the 7th International Symposium of Innovative BioProduction Kobe (iBioK), Jan. 2016, English, International conferenceDynamic metabolic analysis of microorganisms for the development of bio-based chemical productionInvited oral presentation
- 平成27 年度新資源生物変換研究会 シンポジウム, Jan. 2016, Japanese, バイオインダストリー協会, Domestic conference藻類を利用したバイオエタノール、バイオベース化学品生産Oral presentation
- 第11回バイオマス科学会議, Jan. 2016, Japanese, International conferenceリグノセルロース系バイオマスの同時糖化発酵によるフェニル乳酸生産Oral presentation
- The 2015 International Chemical Congress of Pacific Basin Societies (Pacifichem 2015), Dec. 2015, English, International conferenceLive cell imaging and membrane protein mapping with atomic force microscopeOral presentation
- Pacifichem 2015, Dec. 2015, English, International conferenceDynamic metabolic profiling of the marine microalga Chlamydomonas sp. JSC4 and enhancing its oil productionOral presentation
- New Horizon in Biotechnology 2015, Nov. 2015, English, International conferenceDevelopment of cell factories of microalgae and cyanobacteria based on metabolic analysis[Invited]Invited oral presentation
- 藍藻の分子生物学2015, Nov. 2015, Japanese, Domestic conferenceラン藻Arthrospira platenisからの高収率エタノール生産プロセスの開発Oral presentation
- 第67回日本生物工学会大会 国際シンポジウム, Oct. 2015, Japanese, Domestic conference好機・低濃度グルコース条件下におけるSaccharomyces cerevisiaeの転写解析Oral presentation
- 第67回日本生物工学会大会 国際シンポジウム, Oct. 2015, Japanese, Domestic conferenceキチン系基室からのバイオ燃料生産の検討Oral presentation
- 第67回日本生物工学会大会 国際シンポジウム, Oct. 2015, Japanese, Domestic conferenceSaccharomyces cerevisiae の糖代謝における転写制御ネットワークの予測Oral presentation
- The 21th Symposium of Yong Asian Biochemical engineer's Communyty, Oct. 2015, English, International conferenceEffect of PHO13 gene disruption on xylose productivity of yeast harboring xylose isomerase geneOral presentation
- 第67回日本生物工学会大会 国際シンポジウム, Oct. 2015, English, Domestic conferenceDevelopment of microbial cell factories for biorefineryOral presentation
- The 21th Symposium of Yong Asian Biochemical engineer's Communyty, Oct. 2015, English, International conferenceConversion of high phospholipid-containing oils to biodiesel using immobilizes Aspergillus oryzae whole cell biocatalyst ezpressing Fusarium heteroporum lipaseOral presentation
- 第67回日本生物工学会大会 国際シンポジウム, Oct. 2015, Japanese, Domestic conferenceCombined cell-surface display-and secretion-based strategies for production of cellulosic ethanol with Saccharomyces cerevisiaeOral presentation
- 日本植物学会第79回大会, Sep. 2015, Japanese, 新潟コンベンションセンター, Domestic conference藻類のエネルギー生産における物質代謝メカニズムの解析Oral presentation
- iBioK第5回 合成生物工学シンポジウム, Aug. 2015, Japanese, Domestic conferenceメタボロームのターンオーバー解析技術(代謝プロファイリング)の開発と物質生産への応用[Invited]Nominated symposium
- HMTセミナー 物質生産におけるメタボロミクスのいま, Jul. 2015, Japanese, Domestic conferenceマルチオミクス解析に基づく合理的細胞育種技術の開発[Invited]Public discourse
- 11th Annual International Conference of the Metabolomics Society, Jun. 2015, English, Hyatt Regency Burlingame, International conferenceDynamic metabolic profiling of cyanobacteria for bio-based chemical productionOral presentation
- KMB2015, 42nd Annual Meeting & International Symposium, Jun. 2015, English, HICO, Gyeongju, Korea, International conferenceBiofuel Production from Microalgae and Cyanobacteria Based on Metabolic ProfilingOral presentation
- 2015 KSBB Spring Meeting and International Symposium, Apr. 2015, English, International conferenceDevelopment of microbial cell factories based on metabolic engineering[Invited]Invited oral presentation
- 日本農芸化学会2015年度大会, Mar. 2015, Japanese, 日本農芸化学会, 岡山市, Domestic conference産業用酵母のゲノムシャッフリングによる高温及び酸耐性の付与と高効率キシロース発酵Oral presentation
- 日本農芸化学会2015年度大会, Mar. 2015, Japanese, 日本農芸化学会, 岡山市, Domestic conferenceアミラーゼ表層提示酵母によるシアノバクテリアArthrospira platensisからの高濃度エタノール生産Oral presentation
- 広島大学バイオマスイブニングセミナー, Dec. 2014, Japanese, Domestic conference合成生物工学によるバイオ燃料生産に資する微生物の創製[Invited]Public discourse
- 第30回バイオロジカルズ(タンパク医薬)製造技術研究会セミナー, Nov. 2014, Japanese, Domestic conference代謝システム解析に基づく細胞育種とバイオ医薬品生産への応用[Invited]Public discourse
- iBioK 第4回合成生物工学シンポジウム, Nov. 2014, Japanese, Domestic conference代謝システム解析に基づく合理的細胞育種技術の開発Nominated symposium
- バイオマス研究会, Nov. 2014, Japanese, Domestic conference微細藻類の代謝系解析に基づく次世代バイオ燃料生産への挑戦[Invited]Public discourse
- 5th International Symposium on Innovative Bioprocess in Tainan(i-BioT2014), Nov. 2014, English, National Cheng Kung University, Tainan, China, International conferenceThe development of bioconversion process to bio-ethanol from cyanobacteria Spirulina biomassOral presentation
- 5th International Symposium on Innovative Bioprocess in Tainan(i-BioT2014), Nov. 2014, English, National Cheng Kung University, Tainan, China, International conferenceDevelopment of the novel yeast cell-surface display system for consolidated bioethanol production from biomass resourcesOral presentation
- 3rd Asia-Oceania Algae Innovation Summit 2014, Nov. 2014, English, AOAIS2014, 済州、韓国, International conferenceDevelopment of Dynamic Metabolic Profiling of Cyanobacteria and MicroalgaeOral presentation
- 5th International Symposium on Innovative Bioprocess in Tainan(i-BioT2014), Nov. 2014, English, National Cheng Kung University, Tainan, China, International conferenceDevelopment of dynamic metabolic profiling of cyanobacteria and microalgaeOral presentation
- アジレント・メタボロミクスデイズ2014, Oct. 2014, Japanese, Domestic conferenceオミクス解析のバイオリファイナリー[Invited]Public discourse
- Korean Marine Biology and Biotechnology International Symposium, Oct. 2014, English, International conferenceBiodiesel production in marine microalgae through metabolic profiling and an innovative salinity-gradient strategy[Invited]Invited oral presentation
- 日本植物学会第78回大会, Sep. 2014, Japanese, 日本植物学会, 川崎市, Domestic conference代謝系解析に基づくラン藻・微細藻からのバイオ燃料生産への挑戦Oral presentation
- 第66回日本生物工学会大会, Sep. 2014, Japanese, 日本生物工学会, 札幌市, Domestic conference代謝プロファイリングに基づく微生物育種技術の開発と応用Poster presentation
- 第66回日本生物工学会大会, Sep. 2014, Japanese, 日本生物工学会, 札幌市, Domestic conference酵母によるタンパクの高効率細胞表層提示ならびに分泌生産のための新規分泌シグナル配列の検討Poster presentation
- 第66回日本生物工学会大会, Sep. 2014, Japanese, 日本生物工学会, 札幌市, Domestic conference海産性緑藻 Chlamydomonas sp. JSC4 の細胞組成評価と代謝解析に基づいた油脂高生産系の開発Poster presentation
- 第66回日本生物工学会大会, Sep. 2014, Japanese, 日本生物工学会, 札幌市, Domestic conference塩ストレス下での海洋性シアノバクテリアの代謝プロファイリングPoster presentation
- 第66回日本生物工学会大会, Sep. 2014, Japanese, 日本生物工学会, 札幌市, Domestic conferenceバイオ医薬開発に有用なCHO 細胞の改良、培養状態解明につながるデータの構築Poster presentation
- 日本植物学会第78回大会ランチョンセミナー, Sep. 2014, Japanese, 島津製作所, 川崎市, Domestic conferenceバイオリファイナリーに向けた微生物の開発と代謝系解析の応用[Invited]Invited oral presentation
- 第66回日本生物工学会大会, Sep. 2014, Japanese, 日本生物工学会, 札幌市, Domestic conferenceバイオマス糖化液由来成分が大腸菌のフェニル乳酸発酵に与える影響Poster presentation
- 第66回日本生物工学会大会, Sep. 2014, Japanese, 日本生物工学会, 札幌市, Domestic conferenceコリネ菌におけるフルフラールの分解Poster presentation
- 第66回日本生物工学会大会, Sep. 2014, Japanese, 日本生物工学会, 札幌市, Domestic conferenceキシロース資化性酵母における遺伝子発現プロファイルの経時変化解析Poster presentation
- 第66回日本生物工学会大会, Sep. 2014, Japanese, 日本生物工学会, 札幌市, Domestic conferenceアミラーゼ表層提示酵母によるラン藻スピルリナからの高濃度エタノール生産Poster presentation
- 第66回日本生物工学会大会, Sep. 2014, Japanese, 日本生物工学会, 札幌市, Domestic conferencePHO13 遺伝子の欠損がキシロースイソメラーゼ導入酵母のエタノール生産へ与える影響Poster presentation
- 第66回日本生物工学会大会, Sep. 2014, Japanese, 日本生物工学会, 札幌市, Domestic conferenceluyveromyces marxianus の好気条件での糖代謝系の解析Poster presentation
- 第66回日本生物工学会大会, Sep. 2014, English, 日本生物工学会, 札幌市, Domestic conferenceEffect of illumination coupled with nitrogen depletion on biodiesel production of a marine microalga Chlamydomonas sp. JSC4Poster presentation
- 生物工学若手研究者の集い夏のセミナー, Jul. 2014, Japanese, 生物工学会, 神戸市, Domestic conference色素生産を指標としたCoA代謝経路の評価系Poster presentation
- 生物工学若手研究者の集い夏のセミナー, Jul. 2014, Japanese, 生物工学会, 神戸市, Domestic conferenceキシロースイソメラーゼ導入酵母におけるPHO13遺伝子欠損がエタノール生産へ与える影響Poster presentation
- Metabolic Engineering X, Jun. 2014, English, Society for Biological Engineering, Vancouver, Canada, International conferenceDynamic Metabolic Profiling of Cyanobacteria Under Conditions of Nitrate DepletionOral presentation
- 第16回マリンバイオテクノロジー学会大会, May 2014, Japanese, マリンバイオテクノロジー学会, 津市, Domestic conference発酵生産プロセスのための海洋性ラン藻の利用Poster presentation
- 日本化学会第94春季年会, Mar. 2014, Japanese, 日本化学会, 名古屋市, Domestic conference微細藻類によるバイオ燃料や化学物質製造Oral presentation
- 日本農芸化学会2014年度大会, Mar. 2014, Japanese, 日本農芸化学会, 川崎市, Domestic conference統合型リグノセルロース系エタノール生産プロセスに資するセルラーゼ表層提示酵母の開発Oral presentation
- 第16回化学工学会学生発表会(堺大会), Mar. 2014, Japanese, 化学工学会, 大阪市, Domestic conference代謝工学的手法によるグリコーゲン高生産藍藻の創製Oral presentation
- Asian Congress on Biotechnology - ACB 2013, Dec. 2013, English, Asian Federation of Biotechnolgy, New Delhi, India, International conferenceDevelopment of lipid productive engineering with Chlamydomonas sp. Under marine conditionPoster presentation
- ラン藻の分子生物学2013, Nov. 2013, Japanese, かずさDNA研究所, 木更津市, Domestic conferenceラン藻動的代謝プロファイリング技術の開発と応用Oral presentation
- 1st Korea-Japan Microalgae Symposium, Oct. 2013, English, International conferenceDevelopment of microalgal cell factories based on sysmtems biology approach[Invited]Invited oral presentation
- i-BioB 2013, Oct. 2013, English, International conferenceBiofuel production from halophilic algae[Invited]Invited oral presentation
- 第28回つくば藻類・プロティストフォーラム, Oct. 2013, Japanese, Domestic conferencein vivo 13C標識法によるラン藻動的代謝プロファイリング[Invited]Nominated symposium
- INCHEM TOKYO 2013 産学官マッチングフォーラム, Oct. 2013, Japanese, Domestic conference新たな挑戦:イノベーティブ・バイオプロダクション[Invited]Nominated symposium
- 第65回日本生物工学会大会, Sep. 2013, Japanese, 日本生物工学会, 広島市, Domestic conference緑藻Chlamydomonas orbicularisを用いた海水塩存在下での油脂高生産条件の開発Poster presentation
- 第65回日本生物工学会大会, Sep. 2013, Japanese, 日本生物工学会, 広島市, Domestic conference統合型リグノセルロース系エタノール生産プロセスに資する新規酵母細胞表層提示システムの開発Poster presentation
- 第65回日本生物工学会大会, Sep. 2013, Japanese, 日本生物工学会, 広島市, Domestic conference統合バイオプロセスによる環境調和型セルロース系エタノール生産に資する前処理技術の開発Poster presentation
- 第65回日本生物工学会大会, Sep. 2013, Japanese, 日本生物工学会, 広島市, Domestic conference海洋性シアノバクテリアによるバイオリファイナリーのためのグリコーゲン生産Poster presentation
- 第65回日本生物工学会大会, Sep. 2013, Japanese, 日本生物工学会, 広島市, Domestic conference海水中のイオン濃度が糖質源Spirulinaの増殖;光合成に与える影響Poster presentation
- 第65回日本生物工学会大会, Sep. 2013, Japanese, 日本生物工学会, 広島市, Domestic conference稲わら希硫酸処理液のナノフィルトレーションによる糖濃縮Poster presentation
- 第65回日本生物工学会大会, Sep. 2013, Japanese, 日本生物工学会, 広島市, Domestic conferenceキシロース資化性Saccharomyces cerevisiaeにおける代謝プログラム制御によるギ酸;酢酸耐性関連遺伝子の同定Poster presentation
- 第65回日本生物工学会大会, Sep. 2013, Japanese, 日本生物工学会, 広島市, Domestic conferenceキシロース資化性Saccharomyces cerevisiaeにおけるリン酸シグナル伝達系の制御によるキシロースからのエタノール生産の高効率化Poster presentation
- 第65回日本生物工学会大会, Sep. 2013, Japanese, 日本生物工学会, 広島市, Domestic conferencePhototrophic cultivation of a marine microalga Chlamydomonas orbicularis CO2 fixation and biodiesel production:Effect of medium composition, nitrogen deplection, and sea salt concentrationPoster presentation
- 微細藻類研究会2013, Jun. 2013, Japanese, Domestic conferenceメタボローム解析による次世代バイオ燃料生産のための微細藻代謝改変戦略の導出[Invited]Public discourse
- The 3rd International Conference on Algal Biomass、Biofuels and Bioproducts, Jun. 2013, English, Elsevier, Toronto, Canada, International conferenceDirect conversion of cyanobacteria to ethanol without pretreatment or enzymatic hydrolysis processPoster presentation
- 第4回日本光合成学会年会, May 2013, Japanese, 日本光合成学会, 名古屋市, Domestic conferenceシアノバクテリアからの直接バイオエタノール変換技術の開発Poster presentation
- Internatinal meeting of the Microbiorogical Society of Korea, May 2013, English, Microbiorogical Society of Korea, Jeonju, Korea, International conferenceDevelopment of Microbial Cell Factories for Bio-refineryOral presentation
- 日本農芸化学会2013年度大会, Mar. 2013, Japanese, 日本農芸化学会, 仙台市, Domestic conference微細藻バイオリファイナリーに資する動的代謝プロファイリング解析Oral presentation
- 日本農芸化学会2013年度大会, Mar. 2013, Japanese, 日本農芸化学会, 仙台市, Domestic conference発酵阻害物存在下におけるキシロースからのエタノール発酵向上酵母の構築Oral presentation
- 日本農芸化学会2013年度大会, Mar. 2013, Japanese, 日本農芸化学会, 仙台市, Domestic conference耐塩性シアノバクテリアArthrospira platensisからの効率の良いグリコーゲン回収法の構築Oral presentation
- 化学工学会第78年会, Mar. 2013, Japanese, 化学工学会, 豊中市, Domestic conference神戸大学におけるバイオリファイナリーの進展Oral presentation
- 日本農芸化学会2013年度大会, Mar. 2013, Japanese, 日本農芸化学会, 仙台市, Domestic conferenceリグノセルロース系バイオマスからの統合型バイオエタノール生産に資する新規酵母細胞表層提示システムの開発Oral presentation
- SBJ 64th Annual Meeting, Oct. 2012, English, International conferenceBiorefinery from microalgae through fermentation and metabolic engineering[Invited]Invited oral presentation
- 第64回日本生物工学会大会, Oct. 2012, Japanese, 日本生物工学会, 神戸市, Domestic conferenceリパーゼ分泌挙動の異なる糸状菌Rhizopus oryzae のメタボローム解析Oral presentation
- 第64回日本生物工学会大会, Oct. 2012, Japanese, 日本生物工学会, 神戸市, Domestic conferenceイオン液体前処理とキシロース資化性酵母株を用いたバイオマス糖化発酵プロセスの効率化Oral presentation
- 5th International Conference on Industrial Bioprocesses (IFIB-2012), Oct. 2012, English, National Taiwan University of Science and Technology, Taiwan, China, International conferenceDevelopment of sustainable biorefinery based on microalgaeOral presentation
- The 15th International Biotechnology Symposium (IBS 2012), Sep. 2012, English, The Korean Society for Biotechnology and Bioengineering, Deague, Korea, International conferenceFlux balance analysis of xylose-fermenting Saccharomyces cerevisiae strainOral presentation
- The 13th International Congress on Yeasts, Aug. 2012, English, ICY 2012 Organizing Committee, Madison, USA, International conferenceCell recycle batch fermentation of high‐solid lignocellulose using a recombinant cellulase‐displaying yeast strain for high yield ethanol productionOral presentation
- アジレント・メタボロミクスセミナー2012, Jul. 2012, Japanese, アジレント・テクノロジー㈱, 豊中市, Domestic conferenceバイオリファイナリー構築に資する微生物細胞工場の創製Oral presentation
- Metabolic EngineeringⅨ, Jun. 2012, English, Engineering Conferences International, Biarritz, France, International conferenceControl of phosphate metabolism in a xylose-fermenting yeast strain improves ethanol production from xyloseOral presentation
- 4th A-IMBN Annual Conference, Mar. 2012, English, International conferenceMicrobial engineering for production of biofuels and biorefinery products[Invited]Invited oral presentation
- 日本農芸化学会2012年度大会, Mar. 2012, Japanese, 日本農芸化学会, 京都市, Domestic conferenceシアノバクテリアArthrospira platensisを炭素源としたアミラーゼ発現酵母による直接エタノール生産Oral presentation
- 日本農芸化学会2012年度大会, Mar. 2012, Japanese, 日本農芸化学会, 京都市, Domestic conferenceMetabolic pathway engineering in cyanobacteria Synechocystis sp.PCC 6803 for the production of lactateOral presentation
- 第2回イオン液体討論会, Dec. 2011, Japanese, イオン液体研究会, 京都市, Domestic conferenceイオン液体共存下における酵母のキシロース発酵能評価Oral presentation
- 5th International Algae Congress 2011, Dec. 2011, English, DLG BENELUX, Berlin, Germany, International conferenceProduction of bio-ethanol from microalgae through cell surface engineeringOral presentation
- LECOジャパン・食品分析セミナー, Nov. 2011, Japanese, Domestic conference補助食品・スピルリナのメタボローム解析[Invited]Public discourse
- 第31回キャピラリー電気泳動シンポジウム, Nov. 2011, Japanese, 日本分析化学会電気泳動分析研究懇談会, 山形県鶴岡市, Domestic conferenceCE-TOFMSを用いたバイオエタノール生産酵母の代謝プロファイリングOral presentation
- 第63回日本生物工学会大会, Sep. 2011, Japanese, 日本生物工学会, 小金井市, Domestic conference発酵阻害物耐性遺伝子を導入したキシロース資化性二倍体酵母によるリグノセルロース水熱分解液の繰返し発酵Oral presentation
- 第63回日本生物工学会大会, Sep. 2011, Japanese, 日本生物工学会, 小金井市, Domestic conference細胞内酸化還元バランス調整能を付与したキシロース資化性酵母の取得Oral presentation
- 第63回日本生物工学会大会, Sep. 2011, Japanese, 日本生物工学会, 小金井市, Domestic conference回転式発酵槽を用いた稲わら水熱処理物からのエタノール生産の効率化Oral presentation
- 第63回日本生物工学会大会, Sep. 2011, Japanese, 日本生物工学会, 小金井市, Domestic conferenceユーグレナによるバイオ燃料生産のための高密度CO2 固定サイクルの構築Oral presentation
- 第3回 神戸大学バイオサイエンス研究会・若手研究者交流会, Sep. 2011, Japanese, 神戸大学バイオサイエンス研究会・若手研究者交流会, 神戸市, Domestic conferenceメタボロミクスによるリン酸代謝関連遺伝子欠損酵母の発酵特性の解析Poster presentation
- 第63回日本生物工学会大会, Sep. 2011, Japanese, 日本生物工学会, 小金井市, Domestic conferenceフラン化合物存在下でのXR-XDH 発現酵母を用いたキシロース発酵におけるNADH 依存性Adh1過剰発現のインパクトOral presentation
- 第63回日本生物工学会大会, Sep. 2011, Japanese, 日本生物工学会, 小金井市, Domestic conferenceキシロース資化性酵母Saccharomyces cerevisiae におけるPHO13 遺伝子欠損がエタノール生産に与える影響及び発酵阻害物耐性の評価Oral presentation
- 第99回 醗酵学懇話会, Aug. 2011, Japanese, 日本生物工学会 関西支部, 神戸市, Domestic conferenceシンセティックバイオエンジニアリングによるバイオエタノール生産酵母の育種Invited oral presentation
- SIM2011 Annual Meeting and Exhibition, Jul. 2011, English, Society for Industrial Microbiology, New Orleans, USA, International conferenceRepeated batch-fermentation of rice straw hydrolysate to ethanol using metabolically engineered Saccharomyces cerevisiae strainsOral presentation
- SIM2011 Annual Meeting and Exhibition, Jul. 2011, English, Society for Industrial Microbiology, New Orleans, USA, International conferenceDevelopment of microbial cell factories for biofuel production from biomass by conslidated bioprocessingOral presentation
- バイオマスの今、そして未来, Jun. 2011, Japanese, Domestic conference合成生物工学によるバイオ燃料生産のための微生物細胞工場の創製[Invited]Public discourse
- 2011 International Conference for Bioeconomy, Jun. 2011, English, Tianjin Institute of Scientific and Technical Information, Tianjin、China, International conferenceDevelopment of Microbial Cell Factories for Biofuel Production from BiomassOral presentation
- Asian Congress on Biotechnology 2011, May 2011, English, ACB-2011Organizing Committee, Shanghai, China, International conferenceCellulosic ethanol Production in Saccharomyces cerevisiae through Metabolic Engineering Based on Metabolomic ApproachOral presentation
- 日本農芸化学会2011年度大会, Mar. 2011, Japanese, 日本農芸化学会, 京都市, Domestic conferenceセルラーゼ表層提示株を用いた稲わら水熱処理物からの エタノール生産プロセスの効率化Oral presentation
- 日本農芸化学会2011年度大会, Mar. 2011, Japanese, 日本農芸化学会, 京都市, Domestic conferenceセルラーゼを表層提示した高温耐性酵母Kluyveromyces marxianusを用いたセルロースからのエタノール生産Oral presentation
- 日本農芸化学会2011年度大会, Mar. 2011, Japanese, 日本農芸化学会, 京都市, Domestic conferenceキシロース発酵形質転換酵母における代謝コントロールの解析Oral presentation
- 日本農芸化学会2011年度大会, Mar. 2011, Japanese, 日本農芸化学会, 京都市, Domestic conferencePHO13欠損株を用いたキシロースからのエタノール生産Oral presentation
- 化学工学会第15回学生発表会, Mar. 2011, Japanese, (社)化学工学会, 神戸市, Domestic conferencePHO13欠損株を用いたキシロースからのエタノール生産Oral presentation
- 4th Japan-Korea Biomass Symposium, Nov. 2010, English, International conferenceProduction of bio-ethanol from lignocellulosic materials through the engineering of yeast cell surface and metabolic pathway[Invited]Invited oral presentation
- 第62回日本生物工学会大会, Oct. 2010, Japanese, (社)日本生物工学会, 宮崎市, Domestic conference有用希少イノシトールのバイオアベイラビリティーとバイオコンバージョン生産Oral presentation
- 第62回日本生物工学会大会, Oct. 2010, Japanese, (社)日本生物工学会, 宮崎市, Domestic conference微細藻を利用したバイオ燃料生産システムの構築へ向けてOral presentation
- 第62回日本生物工学会大会, Oct. 2010, Japanese, (社)日本生物工学会, 宮崎市, Domestic conference発酵阻害物存在下におけるキシロースからの効率的なエタノール生産Poster presentation
- 第62回日本生物工学会大会, Oct. 2010, Japanese, (社)日本生物工学会, 宮崎市, Domestic conference海洋性微細藻のシステムバイオロジー解析に向けての網羅的代謝プロファイリング技術の開発Poster presentation
- 第62回日本生物工学会大会, Oct. 2010, Japanese, (社)日本生物工学会, 宮崎市, Domestic conferenceワイン酵母OC-2 トリプルマーカー株をプラットフォームとした高β – グルコシダーゼ活性と高キシロシダーゼ活性を持つキシロース発酵酵母の作製Poster presentation
- 第62回日本生物工学会大会, Oct. 2010, Japanese, (社)日本生物工学会, 宮崎市, Domestic conferenceラン藻Spirulina (Arthrospira) platensis のグリコーゲン蓄積量と増殖速度に対する光強度の影響Poster presentation
- 第62回日本生物工学会大会, Oct. 2010, Japanese, (社)日本生物工学会, 宮崎市, Domestic conferenceキシロース資化性形質転換酵母の代謝解析Poster presentation
- 第62回日本生物工学会大会, Oct. 2010, Japanese, (社)日本生物工学会, 宮崎市, Domestic conferenceキシロース資化性Saccharomyces cerevisiae におけるアルコールデヒドロゲナーゼ過剰発現およびフラン化合物存在下でのキシロース発酵特性Poster presentation
- 第5回メタボロームシンポジウム, Sep. 2010, Japanese, メタボロームシンポジウム実行委員会, 山形県鶴岡市, Domestic conferenceメタボロームを利用した新規バイオエタノール生産酵母の育種Oral presentation
- SIM 2010 Annual Meeting, Aug. 2010, English, Society for Industrial Microbiology, San Francisco, USA, International conferenceProduction of bio-ethanol from lignocellulosic materials through engineering yeast cell surface and metabolic pathwayOral presentation
- SIM 2010 Annual Meeting, Aug. 2010, English, Society for Industrial Microbiology, San Francisco, USA, International conferenceEthanol production from cellulosic material using the thermotolerant yeast displaying cellulolytic enzymes on the cell surfaceOral presentation
- The 1st International Symposium on the Nitrogen Nutrition of Plants, Jul. 2010, English, Nitrogen 2010 Organizing Committee, 犬山市, International conferenceDevelopment of bioethanol production from marine microalgaePoster presentation
- 2010 International Symposium on Advanced Biological Engineering, Jul. 2010, English, Biotechnology for Sustainable Industry and Society, Beijing, China, International conferenceDevelopment of bioethanol production from marine microalgaeOral presentation
- The 1st International Symposium on the Nitrogen Nutrition of Plants, Jul. 2010, English, Nitrogen 2010 Organizing Committee, 犬山市, International conferenceAmmonium assimilation in rice leaves: functions of the chloroplastic andcytosolic phosphoenolpyruvate carboxylases (PEPCs) in supplying carbonskeletonsPoster presentation
- Metabolic Engineering VIII: Metabolic Engineering for Green Growth, Jun. 2010, English, Engineering Conferences International, Jeju、Korea, International conferenceMetabolomic approach to identify molecules that are important for acetic acid tolerance in a recombinant xylose-fermenting yeast strainOral presentation
- Microbes at Work JSPS-SLU colloquium, Jun. 2010, English, JSPS-SLU, Uppsala, Sweden, International conferenceEfficient Production of Cellulosic Ethanol in Saccharomyces cerevisiae through Metabolic Engineering Based on a Metabolomic ApproachOral presentation
- RENEWABLE ENERGY 2010, Jun. 2010, English, International Solar Energy Society, 横浜市, International conferenceConstruction of consolidated bioprocesses for production of bio-fuels and chemicals by using cell surface engineered microbial cellsOral presentation
- 日本農芸化学会2010年度大会, Mar. 2010, Japanese, (社)日本農芸化学会, 東京都目黒区, Domestic conference酢酸耐性酵母によるキシロースからの効率的バイオエタノール生産Oral presentation
- 日本農芸化学会2010年度大会, Mar. 2010, Japanese, (社)日本農芸化学会, 東京都目黒区, Domestic conference細胞表層工学技術を用いたヘミセルロース系バイオマスからのエタノール生産Oral presentation
- 日本農芸化学会2010年度大会, Mar. 2010, Japanese, (社)日本農芸化学会, 東京都目黒区, Domestic conferenceバイオマスからの燃料・化学品生産を目指した細胞工場の創製Oral presentation
- 日本農芸化学会2010年度大会, Mar. 2010, Japanese, (社)日本農芸化学会, 東京都目黒区, Domestic conferenceキシロース資化性遺伝子組換え酵母の代謝フラックス解析Oral presentation
- 日本農芸化学会2010年度大会, Mar. 2010, Japanese, (社)日本農芸化学会, 東京都目黒区, Domestic conferenceD-アラビノースは酵母のエリスロアスコルビン酸の内在前駆体かOral presentation
- The 2nd International Symposium of Innovative BioProduction Kobe, Jan. 2010, English, International conferenceA synthetic biology approach for efficient production of cellulosic ethanol in Saccharomyces cerevisiaeInvited oral presentation
- Airbus meeting, Nov. 2009, English, International conferenceEffective biofuel production process from biomass reseources by whole-cell biocatalyst[Invited]Invited oral presentation
- 日本化学会関東支部講演会 「バイオマス変換最前線」, Nov. 2009, Japanese, Domestic conferenceアーミング酵母を用いた統合プロセスによるバイオエタノール生産[Invited]Invited oral presentation
- APBioChEC'09, Nov. 2009, English, APBioChEC'09 Committee, 神戸市, International conferenceDynamic metabolic profiling in plant leaves using in vivo stable isotope labelingOral presentation
- APBioChEC'09, Nov. 2009, English, APBioChEC'09 Committee, 神戸市, International conferenceBioethaol fermentation from mixed sugar by the recombinant yeast with xyloseisomerase pathwayPoster presentation
- APBioChEC'09, Nov. 2009, English, APBioChEC'09 Committee, 神戸市, International conferenceBioethanol production from mixed sugars using sugar uptake ability enhanced yeast strain by overexpression of transportersPoster presentation
- 農芸化学会2009年度大会, Mar. 2009, Japanese, 日本農芸化学会, 福岡県福岡市, Domestic conferenceバイオエタノール生産を目指したスーパー酵母の創製Oral presentation
- 化学工学会第74年会, Mar. 2009, Japanese, 化学工学会, 横浜市, Domestic conferenceキシロースからの効率的なエタノール生産を行う酵母の創製Oral presentation
- NAISTグローバルCOE公開ワークショップ「光合成の環境応答」, Mar. 2009, Japanese, 日本分子生物学会・日本生化学会, 奈良県生駒市, Domestic conferenceC3 photosynthetic pathway analysis by combination of in vivo 13C-labeling and metabolic profiling[Invited]Invited oral presentation
- 日本分子生物学会年会・日本生化学会大会 合同大会, Dec. 2008, Japanese, 日本分子生物学会・日本生化学会, 神戸市, Domestic conference13CO2を用いた同位体標識による葉内代謝物質のターンオーバー解析[Invited]Invited oral presentation
- メタボロームシンポジウム, Oct. 2008, Japanese, 慶應義塾大学先端生命科学研究所, 山形県鶴岡市, Domestic conferencein vivo安定同位体濃縮による葉内炭素代謝ターンオーバー解析Poster presentation
- 関西光合成研究会シンポジウム, Sep. 2008, Japanese, 関西光合成研究会, 神戸市, Domestic conferencein vivo 13Cエンリッチメントによる葉内炭素代謝解析システムの構築[Invited]Invited oral presentation
- 5th International conference on plant metabolomics, Jul. 2008, English, RIKEN Plant Science Center, 横浜市, Domestic conferenceDevelopment of analytical system to monitor metabolic turnoverin plant leaves using in vivo isotope dilution from 13CO2Poster presentation
- 日本農芸化学会大会, Mar. 2008, Japanese, 日本農芸化学会, 名古屋市, Domestic conference葉緑体形質転換技術を用いたアスタキサンチン生産Oral presentation
- 日本農芸化学会大会, Mar. 2008, Japanese, 日本生物工学会, 名古屋市, Domestic conferenceCE/MSを用いた一次代謝物の安定同位体標識率モニタリング法の開発Oral presentation
- 日本植物生理学会年会, Mar. 2008, Japanese, 日本植物生理学会, 札幌市, Domestic conference13CO2エンリッチメントによる代謝ターンオーバー解析Oral presentation
- カロテノイド研究談話会, Sep. 2007, Japanese, 日本カロテノイド研究会, 大阪市, Domestic conference主要カロテノイドとしてアスタキサンチンを生産する葉緑体形質転換植物の作出Oral presentation
- カロテノイド研究談話会, Sep. 2006, Japanese, 日本カロテノイド研究会, 沖縄県, Domestic conference葉緑体形質転換技術を用いたカロテノイド代謝改変植物の作出Oral presentation
- 日本農芸化学会大会, Mar. 2005, Japanese, 日本農芸化学会, 京都市, Domestic conferenceラン藻Synechosystis sp.由来デオキシキシルロースリン酸レダクトイソメラーゼを導入した葉緑体形質転換タバコの作出Oral presentation
- 日本農芸化学会大会, Mar. 2004, Japanese, 日本農芸化学会, 広島市, Domestic conference安定同位体希釈法による定量メタボローム解析法の開発(2)-15N標識による含窒素化合物群の相対定量法-Oral presentation
- 日本生物工学会大会, Sep. 2003, Japanese, 日本生物工学会, 熊本市, Domestic conferenceAspergillus niger由来ペクチンメチルエステラーゼの植物内過剰発現によるメタノール増産と植物矮化Oral presentation
- 日本農芸化学会大会, Mar. 2003, Japanese, 日本農芸化学会, 神奈川県藤沢市, Domestic conferenceアルコールオキシダーゼ・ペルオキシダーゼ二酵素型バイオセンサーを用いたメタノール定量測定Oral presentation
- 日本農芸化学会大会, Mar. 2002, Japanese, 日本農芸化学会, 仙台市, Domestic conferenceAspergillus niger由来ペクチンメチルエステラーゼの植物内過剰発現によるメタノール産生Oral presentation
- 日本生物工学会大会, Aug. 2000, Japanese, 日本生物工学会, 札幌市, Domestic conferenceチューブリン分子に特異的に結合するDNAアプタマーOral presentation
- 学術研究助成基金助成金/基盤研究(C), Apr. 2016 - Mar. 2021Competitive research funding
- 科学研究費補助金/若手研究(A), Apr. 2015 - Mar. 2018, Principal investigatorCompetitive research funding
- 国立研究開発法人科学技術振興機構, 研究成果展開事業 研究成果最適展開支援プログラム(A-STEP)シーズ育成タイプ, 2017, Principal investigator微細藻類からのカロテノイド色素の生産技術開発Competitive research funding
- 三菱ケミカル株式会社, NEDO委託プロジェクト「植物等の生物を用いた高機能品生産技術の開発」, 2017【NEDO:三菱ケミカル】共同研究員受入Competitive research funding
- 一般財団法人日本宇宙フォーラム, 「きぼう」利用フィジビリティスタディテーマ募集, 2017, Principal investigator【JAXA地上】微小重力環境における藻類による物質循環サイクルの実現可能性検証Competitive research funding
- 一般財団法人日本宇宙フォーラム, 「きぼう」利用フィジビリティスタディテーマ募集, 2017, Principal investigator【JAXA装置】微小重力環境における藻類による物質循環サイクルの実現可能性検証Competitive research funding
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