研究者紹介システム

伊藤 洋一郎
イトウ ヨウイチロウ
大学院科学技術イノベーション研究科 科学技術イノベーション専攻
准教授
生物関係
Last Updated :2021/07/25

研究者情報

所属

  • 【主配置】

    大学院科学技術イノベーション研究科 科学技術イノベーション専攻
  • 【配置】

    先端バイオ工学研究センター

学位

  • 博士(学術), 埼玉大学

研究活動

研究分野

  • ナノテク・材料 / 生物分子化学
  • ライフサイエンス / 分子生物学

論文

  • Deletion of DNA ligase IV homolog confers higher gene targeting efficiency on homologous recombination in Komagataella phaffii

    Yoichiro Ito, Toru Watanabe, Shimpei Aikawa, Teruyuki Nishi, Tozo Nishiyama, Yasuyuki Nakamura, Tomohisa Hasunuma, Yuji Okubo, Jun Ishii, Akihiko Kondo

    2018年11月, FEMS Yeast Research, 18 (7), foy074, 英語

    [査読有り]

    研究論文(学術雑誌)

  • A stable, autonomously replicating plasmid vector containing Pichia pastoris centromeric DNA

    Yasuyuki Nakamura, Teruyuki Nishi, Risa Noguchi, Yoichiro Ito, Toru Watanabe, Tozo Nishiyama, Sinpei Aikawa, Tomohisa Hasunuma, Jun Ishii, Yuji Okubo, Akihiko Kondo

    2018年07月, Applied and Environmental Microbiology, 84 (15), e02882 - 17, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Yoichiro Ito, Takao Kitagawa, Mamoru Yamanishi, Satoshi Katahira, Shingo Izawa, Kenji Irie, Makoto Furutani-Seiki, Takashi Matsuyama

    Post-transcriptional upregulation is an effective way to increase the expression of transgenes and thus maximize the yields of target chemicals from metabolically engineered organisms. Refractory elements in the 3' untranslated region (UTR) that increase mRNA half-life might be available. In Saccharomyces cerevisiae, several terminator regions have shown activity in increasing the production of proteins by upstream coding genes; among these terminators the DIT1 terminator has the highest activity. Here, we found in Saccharomyces cerevisiae that two resident trans-acting RNA-binding proteins (Nab6p and Pap1p) enhance the activity of the DIT1 terminator through the cis element GUUCG/U within the 3'-UTR. These two RNA-binding proteins could upregulate a battery of cell-wall-related genes. Mutagenesis of the DIT1 terminator improved its activity by a maximum of 500% of that of the standard PGK1 terminator. Further understanding and improvement of this system will facilitate inexpensive and stable production of complicated organism-derived drugs worldwide.

    NATURE PUBLISHING GROUP, 2016年11月, SCIENTIFIC REPORTS, 6, 36997, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Kentaro Inokuma, Takahiro Bamba, Jun Ishii, Yoichiro Ito, Tomohisa Hasunuma, Akihiko Kondo

    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, 2016年11月, BIOTECHNOLOGY AND BIOENGINEERING, 113 (11), 2358 - 2366, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Yoichiro Ito, Mamoru Yamanishi, Akinori Ikeuchi, Chie Imamura, Takashi Matsuyama

    Combinatorial screening used together with a broad library of gene expression cassettes is expected to produce a powerful tool for the optimization of the simultaneous expression of multiple enzymes. Recently, we proposed a highly tunable protein expression system that utilized multiple genome-integrated target genes to fine-tune enzyme expression in yeast cells. This tunable system included a library of expression cassettes each composed of three gene-expression control elements that in different combinations produced a wide range of protein expression levels. In this study, four gene expression cassettes with graded protein expression levels were applied to the expression of three cellulases: cellobiohydrolase 1, cellobiohydrolase 2, and endoglucanase 2. After combinatorial screening for transgenic yeasts simultaneously secreting these three cellulases, we obtained strains with higher cellulase expressions than a strain harboring three cellulase-expression constructs within one high-performance gene expression cassette. These results show that our method will be of broad use throughout the field of metabolic engineering.

    PUBLIC LIBRARY SCIENCE, 2015年12月, PLOS ONE, 10 (12), e0144870, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Yoichiro Ito, Mamoru Yamanishi, Aldnori Ikeuchi, Takashi Matsuyama

    Control of the expression levels of multiple enzymes in transgenic yeasts is essential for the effective production of complex molecules through fermentation. Here, we propose a tunable strategy for the control of expression levels based on the design of terminator regions and other gene-expression control elements in Saccharomyces cerevisiae. Our genome-integrated system, which is capable of producing high expression levels over a wide dynamic range, will broadly enable metabolic engineering and synthetic biology. We demonstrated that the activities of multiple cellulases and the production of ethanol were doubled in a transgenic yeast constructed with our system compared with those achieved with a standard expression system.

    AMER CHEMICAL SOC, 2015年01月, ACS SYNTHETIC BIOLOGY, 4 (1), 12 - 16, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Yoichiro Ito, Mamoru Yamanishi, Akinori Ikeuchi, Chie Imamura, Kenro Tokuhiro, Takao Kitagawa, Takashi Matsuyama

    Strong terminator regions could be used to improve metabolically engineered yeasts by increasing the target enzyme protein yields above those achieved with traditional terminator regions. We recently identified five strong terminator regions (RPL41Bt, RPL15At, DIT1t, RPL3t, and IDP1t) in a comprehensive analysis of Saccharomyces cerevisiae. The effect of the terminator regions was analyzed by measuring the protein production of a linked transgene, and was shown to be twice that of a traditional terminator region (PGK1t). Here, we investigated whether the activity of the terminator regions is affected by exchange of a strong promoter or reporter in the linked transgene, carbon source for cell growth, stress factors, host yeast strain, or stage of the growth phase. Our results indicate that the activities of all five terminator regions were twice that of PGK1t in all conditions tested. In addition, we demonstrated that the strong activity of these terminator regions could be used to improve secretory production of endoglucanase II derived from Tricodermaressei, and that the DIT1t strain was the best of the five strains for this purpose. We therefore propose that DIT1t, and the four other terminator regions, could be applied to the development of improved metabolically engineered yeasts. (C) 2013 Elsevier B.V. All rights reserved.

    ELSEVIER SCIENCE BV, 2013年12月, JOURNAL OF BIOTECHNOLOGY, 168 (4), 486 - 492, 英語

    [査読有り]

    研究論文(学術雑誌)

  • A Genome-Wide Activity Assessment of Terminator Region in Saccharomyces cervisiae Provides a “Terminatome” Toolbox

    Mamoru Yamanishi, Yoichiro Ito, Reiko Kintaka, Chie Imamura, Satoshi Katahira, Akinori Ikeuchi, Hisao Moriya, Takashi Matsuyama

    2013年02月, ACS Synthetic Biology, 2 (6), 337 - 347, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Y. Ito, A. Ikeuchi, C. Imamura

    We aimed at constructing thermostable cellulase variants of cellobiohydrolase II, derived from the mesophilic fungus Phanerochaete chrysosporium, by using an advanced evolutionary molecular engineering method. By aligning the amino acid sequences of the catalytic domains of five thermophilic fungal CBH2 and PcCBH2 proteins, we identified 45 positions where the PcCBH2 genes differ from the consensus sequence of two to five thermophilic fungal CBH2s. PcCBH2 variants with the consensus mutations were obtained by a cell-free translation system that was chosen for easy evaluation of thermostability. From the small library of consensus mutations, advantageous mutations for improving thermostability were found to occur with much higher frequency relative to a random library. To further improve thermostability, advantageous mutations were accumulated within the wild-type gene. Finally, we obtained the most thermostable variant Mall4, which contained all 15 advantageous mutations found in this study. This variant had the same specific cellulase activity as the wild type and retained sufficient activity at 50°C for > 72 h, whereas wild-type PcCBH2 retained much less activity under the same conditions. The history of the accumulation process indicated that evolution of PcCBH2 toward improved thermostability was ideally and rapidly accomplished through the evolutionary process employed in this study. © 2012 The Author. Published by Oxford University Press. All rights reserved.

    2013年01月, Protein Engineering, Design and Selection, 26 (1), 73 - 79, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Miho Suzuki, Satoshi Tanaka, Yoichiro Ito, Makiko Inoue, Takafumi Sakai, Koichi Nishigaki

    Sensing systems based on Forster resonance energy transfer (FRET) can be used to monitor enzymatic reactions, protein-protein interactions, changes in conformation, and Ca2+ oscillations in studies on cellular dynamics. We developed a series of FRET-based chimeric bioprobes, each consisting of fluorescent protein attached to a fluorescent dye. Green and red fluorescent proteins were used as donors and a series of Alexa Fluor dyes was used as acceptors. The basic fluorescent proteins were substituted with appropriate amino acids for recognition of the target (caspase-3) and subjected to site-directed modification with a fluorescent dye. Variants that retained similar emission profiles to the parent proteins were readily derived for use as FRET-based bioprobes with various fluorescent patterns by incorporating various fluorescent proteins and dyes, the nature of which could be adjusted to experimental requirements. All the constructs prepared functioned as bioprobes for quantitative measurement of caspase-3 activity in vitro. Introduction of the bioprobes into cells was so simple and efficient that activation of caspase-3 upon apoptosis could be monitored by means of cytometric analysis. FRET-based bioprobes are valuable tool for high-throughput flow-cytometric analysis of many cellular events when used in conjunction with other fluorescent labels or markers. Statistical dynamic studies on living cells could provide indications of paracrine signaling. (C) 2011 Elsevier B.V. All rights reserved.

    ELSEVIER SCIENCE BV, 2012年02月, Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1823 (2), 215 - 226, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Yoshihiro Shimizu, Saburo Tsuru, Yoichiro Ito, Bei-Wen Ying, Tetsuya Yomo

    Population adaptation can be determined by stochastic switching in living cells. To examine how stochastic switching contributes to the fate decision for a population under severe stress, we constructed an Escherichia coli strain crucially dependent on the expression of a rewired gene. The gene essential for tryptophan biosynthesis, trpC, was removed from the native regulatory unit, the Trp operon, and placed under the extraneous control of the lactose utilisation network. Bistability of the network provided the cells two discrete phenotypes: the induced and suppressed level of trpC. The two phenotypes permitted the cells to grow or not, respectively, under conditions of tryptophan depletion. We found that stochastic switching between the two states allowed the initially suppressed cells to form a new population with induced trpC in response to tryptophan starvation. However, the frequency of the transition from suppressed to induced state dropped off dramatically in the starved population, in comparison to that in the nourished population. This reduced switching rate was compensated by increasing the initial population size, which probably provided the cell population more chances to wait for the rarely appearing fit cells from the unfit cells. Taken together, adaptation of a starved bacterial population because of stochasticity in the gene rewired from the ancient regulon was experimentally confirmed, and the nutritional status and the population size played a great role in stochastic adaptation.

    PUBLIC LIBRARY SCIENCE, 2011年09月, PLOS ONE, 6 (9), e23953, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Yuki Matsumoto, Yoichiro Ito, Saburo Tsuru, Bei-Wen Ying, Tetsuya Yomo

    A synthetic dual-function operon with a bistable structure was designed and successfully integrated into the bacterial genome. Bistability was generated by the mutual inhibitory structure comprised of the promoters P-tet and P-lac and the repressors LacI and TetR. Dual function essential for cell growth was introduced by replacing the genes (i.e., hisC and leuB) encoding proteins involved in the biosynthesis of histidine and leucine from their native chromosomal locations to the synthetic operon. Both colony formation and population dynamics of the cells carrying this operon showed that the cells survived starvation and the newly formed population transited between the two stable states, representing the induced hisC and leuB levels, in accordance with the nutritional status. The results strongly suggested that the synthetic design of proto-operons sensitive to external perturbations is practical and functional in native cells.

    HINDAWI PUBLISHING CORPORATION, 2011年, JOURNAL OF BIOMEDICINE AND BIOTECHNOLOGY, 489265, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Bei-Wen Ying, Yoichiro Ito, Yoshihiro Shimizu, Tetsuya Yomo

    Genetic reconstruction of regulatory gene circuits is currently applied in systematic dynamics and structure-function studies of intact cellular networks in systems biology. We present a modified procedure for the integration of complex genetic circuits into the Escherichia coli genome, to provide an efficient synthetic approach for stochastic study and the artificial engineering of genetic networks. Linear artificial sequences of various lengths were easily integrated into the bacterial genome at one time. Comparison of the cellular concentrations of proteins encoded by genes carried on plasmids or the genome indicated that genome recombination could minimize the copy number noise in the genetic circuit, allowing precise design and interpretation of the cellular network. The refined recombination procedure allowed efficient construction of a single copy of a complex genetic circuit in cells, and the resultant reduced fluctuation in copy number led to accurate phenotypic behaviour of the genome-integrated synthetic switch corresponding to the design principle. The availability of long-fragment insertions makes the reconstruction of complex networks easy on the genome, and provides a powerful tool for precise engineering in synthetic and systems biology. (C) 2010, The Society for Biotechnology, Japan. All rights reserved.

    SOC BIOSCIENCE BIOENGINEERING JAPAN, 2010年11月, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 110 (5), 529 - 536, 英語

    [査読有り]

    研究論文(学術雑誌)

  • How selection affects phenotypic fluctuation

    Yoichiro Ito, Hitoshi Toyota, Kunihiko Kaneko, Tetsuya Yomo

    2009年04月, Molecular Systems Biology, 5, 264, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Tetsuya Yomo, Katsuhiko Sato, Yoichiro Ito, Kunihiko Kaneko

    Springer, 2006年, Biologically Inspired Approaches to Advanced Information Technology, Second International Workshop, BioADIT 2006, Osaka, Japan, January 26-27, 2006, Proceedings, 107 - 112

    [査読有り]

  • M Suzuki, Y Ito, Sakata, I, T Sakai, Y Husimi, KT Douglas

    Green fluorescent protein (UV5) was re-engineered to remove native cysteine residues, and a new cysteine was introduced near the C-terminus, similar to 20 angstrom from the native fluorophore, for site-specific attachment of chemical fluorophores. The resultant efficient intramolecular FRET quenched GFP emission and gave a new emission band from the conjugated fluorophore. Caspase-3 cleavage of constructs with a caspase-3 sequence near the C-terminus in the sequence between the native fluorophore and the new cysteine, located C-terminal to the caspase site, destroyed the FRET, the emitted color reverting to that of unmodified GFP. This process was demonstrated in vitro with caspase-3 and lysates from cells undergoing apoptosis. Real-time emission changes for the Alexa Fluor 532 conjugate of this GFP, studied quantitatively in vivo for single HeLa cells using the ratios of fluorescence at the red and green maxima by confocal microscopy, showed that caspase-3 action in the cytosol preceded that in the nucleus. (c) 2005 Elsevier Inc. All rights reserved.

    ACADEMIC PRESS INC ELSEVIER SCIENCE, 2005年05月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 330 (2), 454 - 460, 英語

    [査読有り]

    研究論文(学術雑誌)

  • A Chemically Modified Green-Fluorescent Protein that Responds to Cleavage of an Engineered Disulphide Bond by Fluorescence Resonance Energy Transfer (FRET)-Based Changes

    Wei Zhang, Miho Suzuki, Yoichiro Ito, Kenneth T. Douglas

    2005年04月, Chemistry Letters, 34 (6), 766 - 767, 英語

    [査読有り]

    研究論文(学術雑誌)

  • M Suzuki, Y Ito, HE Savage, Y Husimi, KT Douglas

    The native cysteine residues of green fluorescent protein (GFP) at positions 48 and 70 were replaced by non-thiolic amino acids, and new cysteine sites were introduced at specific, surface positions. Based on molecular modeling of the GFP structure, the sites chosen for mutagenesis to Cys were glutamic acid at position 6 and isoleucine at position 229. These new, unique cysteine sites provided reactive thiol groups suitable for site-specific chemical modification by eosin-based fluorescence labels. The new constructs were designed to serve as the basis of proof of principle for fluorescence resonance energy transfer (FRET) using an enzyme-activated (trypsin) intervening sequence between native and chemically conjugated fluorophores. These eosin moieties provided chemical FRET partners for the native GFP chromophore. On excitation, these GFP-eosin constructs exhibited strong intramolecular FRET, with quenching of the native GFP (511 nm) fluorophore emission and emission around 540 nm, corresponding to eosin. GFP mutants engineered with trypsin-sensitive sequences close to the eosin site, so that on trypsinolysis FRET was destroyed, the emission wavelength switching from that of the chemical FRET partner back to that of the native GFP fluorophore, providing efficient, ratio-based detection. This protein engineering provides the basis for novel bioprobes for enzymatic triggering using intramolecular FRET between GFP and carefully sited chemical labels. (C) 2004 Elsevier B.V. All rights reserved.

    ELSEVIER SCIENCE BV, 2004年09月, BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION, 1679 (3), 222 - 229, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Y Ito, T Kawama, Urabe, I, T Yomo

    We have investigated the evolvability of an insoluble random polypeptide, RP3-34, to a soluble form through iterative mutation and selection with the aid of the green fluorescent protein (GFP) folding reporter. To assess the solubility of the polypeptides in the selected clones of each generation, the polypeptide genes were detached from the GFP fusions and expressed with a His(6) tag. The solubility of the variant random polypeptides increased in each generation within the scope of the evolutionary process, and the polypeptides assumed a soluble form from the fourth generation. Analysis of the synonymous and nonsynonymous mutations found in the deduced amino acid sequence of the selected polypeptides revealed that selection had accelerated the evolutionary rate. The solubility and hydrophobicity of the polypeptides and the 25 arbitrarily chosen random polypeptides found in a previously prepared library were determined, analyzed, and interpreted from the landscape on the protein sequence space. This study showed the evolvability of an insoluble arbitrary sequence toward a soluble one, hence, it provides a new perspective on the field of artificial evolution.

    SPRINGER-VERLAG, 2004年02月, JOURNAL OF MOLECULAR EVOLUTION, 58 (2), 196 - 202, 英語

    [査読有り]

    研究論文(学術雑誌)

  • K Sato, Y Ito, T Yomo, K Kaneko

    A general relationship between fluctuation and response in a biological system is presented. The fluctuation is given by the variance of some quantity, whereas the response is given as the average change of that quantity for a given parameter change. We propose a relationship where the two are proportional, in a similar way to the fluctuation-dissipation theorem in physics. By studying an evolution experiment where fluorescence of protein in bacteria increases, we confirm our relation by observing a positive correlation between the speed of fluorescence evolution and the phenotypic fluctuation of the fluorescence over clone bacteria. The generality of the relationship as well as its relevance to evolution is discussed.

    NATL ACAD SCIENCES, 2003年11月, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 100 (24), 14086 - 14090, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Intramolecular Fluorescent Resonance Energy Transfer (FRET) by BODIPY Chemical Modification of Cysteine-engineered Mutants of Green Fluorescent Protein

    Miho Suzuki, Yoichiro Ito, Elizabeth Savage Hannah, Yuzuru Husimi, Kenneth T. Douglas

    2003年02月, Chemistry Letters, 32 (3), 306 - 307, 英語

    [査読有り]

    研究論文(学術雑誌)

  • Miho Suzuki, Yoichiro Ito, Hannah Elizabeth Savage, Yuzuru Husimi, Kenneth T. Douglas

    Specific, surface cysteine sites have been introduced into Green Fluorescent Protein (GFP) to allow site-specific chemical modification by thiol-directed reagents. These sites have been labelled using BODIPY/eosin/rhodamine reagents as chemical FRET partners for the native GFP chromophore. When they were excited at 488 nm these engineered GFP: conjugated-fluorophore constructs, showed quenching of the native GFP fluorophore emission at 511 nm. New emission bands appeared corresponding to each chemical fluorophore emission. Thus the new GFP chimeras exhibited strong intramolecular FRET. GFP mutants were then engineered with trypsin-sensitive sequences located close to the chemical fluorophore-bearing cysteine site. Trypsinolysis caused major changes in the FRET fluorescence spectra. On trypsinolysis the FRET was destroyed, as the FRET partners were now on separate molecules in the cleaved products. T Consequently, the emission wavelength altered from that of the chemically conjugated FRET partner back to that of the native fluorophore of the GFP (511 nm). This provides the possibility of efficient, ratio-based detection. Thus, protein re-engineering has led to novel probes capable of enzymatic triggering based on intramolecular FRET between GFP and specifically sited chemical labels.

    2003年, Proceedings of SPIE - The International Society for Optical Engineering, 4967, 1 - 10, 英語

    [査読有り]

    研究論文(国際会議プロシーディングス)

  • A T-extended vector using a green fluorescent protein as an indicator

    Y Ito, M Suzuki, Y Husimi

    T-extended vector (T-vector) is a useful tool for cloning PCR products directly. We exploited a novel T-vector using a green fluorescent protein (GFP) as an indicator based on insertional inactivation. The brightest GFP mutant was used for easy detection even under daylight. The 100 bp and 0.9 kb of PCR products were cloned, and the transformant colonies with inserts were adjudged by the fluorescent green-white screening. The GFP system was more sensitive to insertional inactivation than the P-galactosidase system at the conventional insertion sites. (C) 2000 Published by Elsevier Science B.V. All rights reserved.

    ELSEVIER SCIENCE BV, 2000年03月, GENE, 245 (1), 59 - 63, 英語

    [査読有り]

    研究論文(学術雑誌)

  • A novel mutant of green fluorescent protein with enhanced sensitivity for microanalysis at 488 nm excitation

    Y Ito, M Suzuki, Y Husimi

    Green fluorescent protein (GFP) has been utilized as a powerful reporter of gene expression and protein localization in cells. We discovered a mutant carrying point mutation S208L from a UV-excitable GFP (F99S/M153T/V163A). It had the enhanced fluorescence intensity. Introduction of the red-shifted mutations (F64L/S65T) to this mutant led to the GFP having the brightest mutants reported which were expressed in Escherichia coli and excited at 488 nm. The relative fluorescence intensities to that of wild-type GFP and GFPuv were increased about 120- and 10-fold, respectively. It was shown that the S208L mutation contributes to both a higher intrinsic brightness of GFP and a higher expression level in E. coli, (C) 1999 Academic Press.

    ACADEMIC PRESS INC, 1999年10月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 264 (2), 556 - 560, 英語

    [査読有り]

    研究論文(学術雑誌)

MISC

  • 低分子抗体医薬の開発展望

    中村 泰之, 伊藤 洋一郎, 梅津 光央, 石井 純, 近藤 昭彦

    シーエムシー出版, 2017年06月, 月刊BIO INDUSTRY, 34 (6), 54 - 62, 日本語

    [招待有り]

    記事・総説・解説・論説等(商業誌、新聞、ウェブメディア)

  • A genome-wide activity assessment of terminator regions in Saccharomyces cerevisiae provides a "terminatome" toolbox

    Yoichiro Ito, Mamoru Yamanishi, Takashi Matsuyama

    WILEY-BLACKWELL, 2013年09月, YEAST, 30, 91 - 91, 英語

    研究発表ペーパー・要旨(国際会議)

講演・口頭発表等

  • Pichia 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

    伊藤 洋一郎, 石上 美佐, 橋場 倫子, 中村 泰之, 蓮沼 誠久, 石井 純, 近藤 昭彦

    第71回日本生物工学会大会, 2019年09月, 日本語, 岡山大学 津島キャンパス, 国内会議

    口頭発表(一般)

  • 分泌タンパク質精製のハイスループット自動化

    堀川 拓真, 伊藤 洋一郎, 石井 純, 近藤 昭彦

    生物工学若手研究者の集い 夏のセミナー2019, 2019年07月, 日本語, 琵琶湖国定公園 近江白浜 政府登録旅館 白浜荘, 国内会議

    ポスター発表

  • 抗体生産性に関わる遺伝子探索のためのPichia pastorisゲノム編集技術の構築

    山田 健人, 南部-西田, 由美子, 伊藤 洋一郎, 西田 敬二, 石井 純, 近藤 昭彦

    生物工学若手研究者の集い 夏のセミナー2019, 2019年07月, 日本語, 琵琶湖国定公園 近江白浜 政府登録旅館 白浜荘, 国内会議

    ポスター発表

  • Construction of a stable, autonomously replicating plasmid vector containing Pichia pastoris centromeric DNA

    Yasuyuki Nakamura, Teruyuki Nishi, Risa Noguchi, Yoichiro Ito, Toru Watanabe, Tozo Nishiyama, Shimpei Aikawa, Tomohisa Hasunuma, Jun Ishii, Yoshihiko Yasohara, Akihiko Kondo

    The Symposium on Biorefinery and Biprocess Topics, 2018 (iBio-N 2018), 2018年11月, 英語, Nanjing, China, 国際会議

    ポスター発表

  • 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

    伊藤 洋一郎, 渡邉 徹, 藍川 晋平, 西 輝之, 西山 陶三, 中村 泰之, 蓮沼 誠久, 八十原 良彦, 石井 純, 近藤 昭彦

    第70回日本生物工学会大会, 2018年09月, 日本語, 日本生物工学会, 関西大学 千里山キャンパス, 国内会議

    口頭発表(一般)

  • Pichia pastorisにおける自律複製型ベクターを用いた効率的なDNAマルチアセンブル法

    西 輝之, 西山 陶三, 山路 大樹, 玉井 雅也, 渡邉 徹, 中村 泰之, 伊藤 洋一郎, 石井 純, 近藤 昭彦, 八十原 良彦

    第70回日本生物工学会大会, 2018年09月, 日本語, 日本生物工学会, 関西大学 千里山キャンパス, 国内会議

    口頭発表(一般)

  • Pichia 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

    中村 泰之, 西 輝之, 野口 理紗, 伊藤 洋一郎, 渡邉 徹, 西山 陶三, 藍川 晋平, 蓮沼 誠久, 石井 純, 八十原 良彦, 近藤 昭彦

    第70回日本生物工学会大会, 2018年09月, 日本語, 日本生物工学会, 関西大学 千里山キャンパス, 国内会議

    口頭発表(一般)

  • P. pastorisにおける自律複製型ベクターを用いた効率的なDNAマルチアセンブル法

    西 輝之, 山路 大樹, 玉井 雅也, 渡邉 徹, 西山 陶三, 中村 泰之, 伊藤 洋一郎, 石井 純, 近藤 昭彦, 八十原 良彦

    第12回日本ゲノム微生物学会年会, 2018年03月, 日本語, 日本ゲノム微生物学会, 京都大学 桂キャンパス, 国内会議

    口頭発表(一般)

  • Enhanced protein secretion by accumulation of novel effective factors in Pichia pastoris

    Yoichiro Ito, Yasuyuki Nakamura, Shimpei Aikawa, Tomohisa Hasunuma, Jun Ishii, Akihiko Kondo

    The 9th International Symposium of Innovative BioProduction Kobe (iBioK), 2018年02月, 英語, Kobe, Japan, 国際会議

    ポスター発表

  • A new autonomous replicating plasmid vector containing Pichia pastoris centromeric DNA

    Yasuyuki Nakamura, Teruyuki Nishi, Risa Noguchi, Yoichiro Ito, Toru Watanabe, Tozo Nishiyama, Shimpei Aikawa, Tomohisa Hasunuma, Jun Ishii, Yuji Okubo, Akihiko Kondo

    The 9th International Symposium of Innovative BioProduction Kobe (iBioK), 2018年02月, 英語, Kobe, Japan, 国際会議

    ポスター発表

  • Pichia 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

    伊藤 洋一郎, 中村 泰之, 西 輝之, 藍川 晋平, 蓮沼 誠久, 石井 純, 近藤 昭彦

    第69回日本生物工学会大会, 2017年09月, 日本語, 日本生物工学会, 早稲田大学 西早稲田キャンパス, 国内会議

    ポスター発表

  • Pichia 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

    中村 泰之, 西 輝之, 野口 理紗, 伊藤 洋一郎, 渡邉 徹, 西山 陶三, 藍川 晋平, 蓮沼 誠久, 石井 純, 八十原 良彦, 近藤 昭彦

    第69回日本生物工学会大会, 2017年09月, 日本語, 日本生物工学会, 早稲田大学 西早稲田キャンパス, 国内会議

    ポスター発表

  • Enhanced protein secretion by accumulation of novel effective factors in Pichia pastoris

    Yoichiro Ito, Yasuyuki Nakamura, Shimpei Aikawa, Tomohisa Hasunuma, Jun Ishii, Akihiko Kondo

    The 14th International Congress on Yeasts (ICY2016), 2016年09月, 英語, ICY14組織委員会, 淡路市, 国際会議

    ポスター発表

  • A new autonomous replicating plasmid vector containing centromere DNA sequence of Pichia pastoris

    Yasuyuki Nakamura, Teruyuki Nishi, Risa Noguchi, Yoichiro Ito, Toru Watanabe, Tozo Nishiyama, Shimpei Aikawa, Tomohisa Hasunuma, Jun Ishii, Yoshihiko Yasohara, Akihiko Kondo

    The 14th International Congress on Yeasts (ICY2016), 2016年09月, 英語, Awaji, Japan, 国際会議

    ポスター発表