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浅水 俊平
先端バイオ工学研究センター
准教授

研究者基本情報

■ 学位
  • 博士（工学）, 富山県立大学
■ 研究キーワード
  • 放線菌
  • 二次代謝経路
  • 天然物生合成
  • 微生物間相互作用
  • ゲノムマイニング
  • 安定同位体ラベリング
  • 比較メタボローム解析
  • 比較トランスクリプトーム解析
  • 抗生物質
  • トロイの木馬抗生物質
  • シデロフォア
  • リボソーム翻訳後修飾ペプチド
  • リポペプチド
  • アミノシクリトール
  • 細菌アルカロイド
■ 研究分野
  • ライフサイエンス / 応用微生物学
  • ライフサイエンス / 応用生物化学
■ 委員歴
  • 2022年08月 - 現在, Frontiers in Microbiology (Microbial Physiology and Metabolism), Associate Editor

研究活動情報

■ 受賞
  • 2023年03月 日本農芸化学会, 日本農芸化学会2023年度広島大会（第13回）トピックス賞, 放線菌におけるヒ素二次代謝経路に関する研究

  • 2022年03月 日本農芸化学会, 日本農芸化学会2022年度京都大会（第12回）トピックス賞, 土壌微生物と共存する新規ウイルス様粒子の生物学的意義

  • 2021年03月 日本農芸化学会, 日本農芸化学会2021年度仙台大会（第11回）トピックス賞, goadvioninにおける脂肪酸部位の生合成の解析

  • 2018年03月 日本農芸化学会, 日本農芸化学会2018年度名古屋大会（第9回）トピックス賞, 新規labionen構造の形成に関与するlanthipeptide合成酵素の解析

  • 2016年09月 日本放線菌学会, 浜田賞, 放線菌の窒素含有天然物生合成に関する研究

  • 2016年03月 日本農芸化学会, 農芸化学奨励賞, 放線菌由来窒素含有天然生物活性物質の生合成に関する研究

■ 論文
  • Fumihiro Ishikawa, Maya Nohara, Akimasa Miyanaga, Satoki Kuramoto, Natsuki Miyano, Shumpei Asamizu, Fumitaka Kudo, Hiroyasu Onaka, Tadashi Eguchi, Genzoh Tanabe
    Nonribosomal peptides (NRPs), one of the most widespread secondary metabolites in nature, with therapeutically significant activities, are biosynthesized by modular nonribosomal peptide synthetases (NRPSs). Aryl acids contribute to the structural diversity of NRPs as well as nonproteinogenic amino acids and keto acids. We previously confirmed that a single Asn-to-Gly substitution in the 2,3-dihydroxybenzoic acid-activating adenylation (A) domain EntE involved in enterobactin biosynthesis accepts monosubstituted benzoic acid derivatives with nitro, cyano, bromo, and iodo functionalities at the 2 or 3 positions. Here, we showed that the mutant EntE (N235G) accommodates various disubstituted benzoic acid derivatives with halogen, methyl, methoxy, nitro, and cyano functionalities at the 2 and 3 positions and monosubstituted benzoic acid with an alkyne at the 3 position. Structural analysis of the mutant EntE (N235G) with nonhydrolyzable aryl-AMP analogues using 3-chloro-2-methylbenzoic acid and 3-prop-2-ynoxybenzoic acid revealed how bulky 3-chloro-2-methylbenzoic acid and clickable 3-prop-2-ynoxybenzoic acid are recognized by enlarging the substrate-binding pocket of the enzyme. When engineered EntE mutants were coupled with enterobactin and vibriobactin biosynthetic enzymes, 3-hydroxybenzoic acid-, salicylic acid-, and 3-bromo-2-fluorobenzoic acid-containing peptides were produced as early stage intermediates, highlighting the potential of NRP biosynthetic pathway engineering for constructing diverse aryl acid-containing metabolites.
    2024年12月, ACS chemical biology, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Yukun Lei, Hiroyasu Onaka, Shumpei Asamizu
    Combined-cultures involving mycolic acid-containing bacteria (MACB) can stimulate secondary metabolite (SM) production in actinomycetes. In a prior investigation, we screened Streptomyces coelicolor JCM4020 mutants with diminished production of SMs, specifically undecylprodigiosin (RED), which was enhanced by introducing the MACB Tsukamurella pulmonis TP-B0596. In this study, we conducted mutational analysis that pinpointed the sco1842 gene, which we assigned the gene name ccr1 (combined-culture related regulatory protein no. 1), as a crucial factor in the deficient phenotype observed in the production of various major SMs in S. coelicolor A3(2). Notably, the Ccr1 (SCO1842) homolog was found to be highly conserved throughout the Streptomyces genome. Although Ccr1 lacked conserved motifs, in-depth examination revealed the presence of a helix-turn-helix (HTH) motif in the N-terminal region and a helicase C-terminal domain (HCTD) motif in the C-terminal region in some of its homologs. Ccr1 was predicted to be a nucleoid-associated protein (NAP), and its impact on gene transcription was validated by RNA-seq analysis that revealed genome-wide variations. Furthermore, RT-qPCR demonstrated that ccr1 was transcriptionally activated in combined-culture with T. pulmonis, which indicated that Ccr1 is involved in the response to bacterial interaction. We then investigated Streptomyces nigrescens HEK616 in combined-culture, and the knockout mutant of the ccr1 homolog displayed reduced production of streptoaminals and 5aTHQs. This finding reveals that the Ccr1 homolog in Streptomyces species is associated with SM production. Our study elucidates the existence of a new family of NAP-like proteins that evolved in Streptomyces species and play a pivotal role in SM production.
    2024年07月, Frontiers in microbiology, 15, 1422977 - 1422977, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）
    神戸大学リポジトリ（Kernel）へのリンク

  • Kimberly S. Taylor, Madison M. McMonagle, Schaelee C. Guy, Ariana M. Human-McKinnon, Shumpei Asamizu, Heidi J. Fletcher, Bradley W. Davis, Takashi L. Suyama
    An organo–ruthenium catalyst conjugated to albumin efficiently unmasks an alloc group under physiologically relevant conditions.
    Royal Society of Chemistry (RSC), 2024年04月, Organic & Biomolecular Chemistry, 22(15) (15), 2992 - 3000, 英語
    [査読有り]
    研究論文（学術雑誌）

  • Shotaro Hoshino, Shinta Ijichi, Shumpei Asamizu, Hiroyasu Onaka
    American Chemical Society (ACS), 2023年08月, Journal of the American Chemical Society, 145(32) (32), 17863 - 17871, 英語
    [査読有り]
    研究論文（学術雑誌）

  • Fumihiro Ishikawa, Natsumi Tsukumo, Erika Morishita, Shumpei Asamizu, Saaya Kusuhara, Shinsuke Marumoto, Katsuki Takashima, Hiroyasu Onaka, Genzoh Tanabe
    Coupled with precursor-directed biosynthesis, activity-based protein profiling of non-ribosomal peptide synthetases provides rational guidance for the biosynthetic diversification of non-ribosomal peptides.
    Royal Society of Chemistry (RSC), 2023年08月, Chemical Communications, 59(62) (62), 9473 - 9476, 英語
    [査読有り]
    研究論文（学術雑誌）

  • Toshiki Nagakubo, Shumpei Asamizu, Tatsuya Yamamoto, Manami Kato, Tatsuya Nishiyama, Masanori Toyofuku, Nobuhiko Nomura, Hiroyasu Onaka
    Contractile injection systems (CISs) are a large group of phage tail-like nanostructures conserved among bacteria. Despite their wide distribution, the biological significance of CISs in bacteria remains largely unclear except for a few unicellular bacteria. Here, we show that Streptomyces lividans-a model organism of filamentous Gram-positive bacteria with highly conserved CIS-related gene clusters-produces intracellular CIS-like nanostructures (Streptomyces phage tail-like particles [SLPs]) that affect phenotypes of this bacterium under hyperosmotic conditions. In contrast to typical CISs released from the cells, SLPs are localized in the cytoplasm of S. lividans. In addition, loss of SLPs leads to (i) delayed erection of aerial mycelia on hyperosmotic solid medium and (ii) decreased growth during the transition from exponential growth phase to stationary phase in hyperosmotic liquid medium. Localization of fluorescent protein-tagged SLPs showed partial correlation with cell wall synthesis-related proteins, including MreB, an actin-like cytoskeleton protein. Our pulldown assay and subsequent quantitative proteome analysis also suggest that 30S ribosomal proteins and cell wall-related proteins, including MreB, are coeluted with SLPs. Furthermore, an interaction assay using the recombinant proteins revealed a direct interaction between a sheath protein of SLP and ribosomal protein S16. Results of cross-linking experiments show indirect interactions between SLPs and translation elongation factors. These findings collectively suggest that SLPs are directly or indirectly associated with a protein interaction network within the cytoplasm of S. lividans and that SLP loss ultimately affects the susceptibility of the bacterium to certain stress conditions. IMPORTANCE Recent bioinformatic analyses have revealed that CIS-related gene clusters are highly conserved in Gram-positive actinomycetes, especially members of the genus Streptomyces known for their ability to produce therapeutic antibiotics. While typical CISs are released from the cells and can act as protein translocation systems that inject effector proteins into the target cells, our results indicate the unique intracellular localization of SLPs, CIS-related nanostructures produced by S. lividans. In addition, the direct and indirect interactions of SLPs with cytoplasmic proteins and SLP localization within specific regions of mycelia suggest that the biological significance of SLPs is related to intracellular processes. Further, SLP loss leads to increased susceptibility of S. lividans to osmotic stress, suggesting that production of these phage tail-like nanostructures ultimately affects the fitness of the bacterium under certain stress conditions. This work will provide new insight into the phage tail-like nanostructures highly conserved in Streptomyces species.
    2023年06月, mSphere, 8(3) (3), e0011423, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Shinta Ijichi, Shotaro Hoshino, Shumpei Asamizu, Hiroyasu Onaka
    Ribosomally synthesized and posttranslationally modified peptides (RiPPs) with polar-functionalized fatty acyl groups are newly found lipopeptide-class natural products. We recently employed a combined approach of genome mining and stable isotope labeling and discovered solabiomycins as one of the polar-functionalized fatty-acylated RiPPs (PFARs) from Streptomyces lydicus NBRC13058. The solabiomycins contained a characteristic sulfoxide group in the labionin moiety referred to as the 'solabionin' structure for the RiPP moiety. A previous gene knockout experiment indicated that solS, which encodes a putative flavin adenine dinucleotide (FAD)-nicotinamide adenine dinucleotide (phosphate) (NAD(P))-binding protein, is involved in the sulfoxidation of an alkyl sulfide in the solabionin. In this study, we isolated deoxysolabiomycins A and B from ΔsolS mutant and fully determined the chemical structures using a series of NMR experiments. We also tested the bioactivity of deoxysolabiomycins against Gram-positive bacteria, including Mycolicibacterium smegmatis, and notably found that the sulfoxide is critical for the antibacterial activity. To characterize the catalytic activity of SolS, the recombinant protein was incubated with a putative substrate, deoxysolabiomycins, and the cofactors FAD and NADPH. In vitro reactions demonstrated that SolS catalyzes the sulfoxidation, converting deoxysolabiomycins to solabiomycins.
    2023年06月, Bioorganic & Medicinal Chemistry Letters, 89, 129323, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Yukun Lei, Shumpei Asamizu, Takumi Ishizuka, Hiroyasu Onaka
    Streptomyces spp. are well-known producers of bioactive secondary metabolites (SMs) that serve as pharmaceutical agents. In addition to their ability to produce SMs, Streptomyces spp. have evolved diverse membrane transport systems to protect cells against antibiotics produced by itself or other microorganisms. We previously screened mutants of Streptomyces coelicolor that show a phenotype of reduced undecylprodigiosin (RED) production in a combined-culture with Tsukamurella pulmonis. Here, we identified a point mutation, which reduced RED production, by performing genome resequencing and genetic complementation. We found that inactivation of the sco1718 gene encoding the TetR family transcriptional regulator (TFR) produced a deficient phenotype for several SMs in Streptomyces coelicolor A3(2). In the genome of S. coelicolor A3(2), two other sets of TFR and two-component ATP-binding cassette (ABC) transporter genes (sco4358-4360 and sco5384-5382) were found which had similar effects on the phenotype for both secondary metabolism and antibiotic resistance. An electrophoretic mobility shift assay and quantitative reverse transcription-PCR experiments demonstrated that TFRs repressed the expression of each adjacent two-component ABC transporter genes by binding to the operator sequence. Notably, the Δsco1718 mutant showed increased resistance to several antibiotics of other actinomycete origin. Our results imply the switching of cell metabolism to direct offense (antibiotic production) or defense (efflux pump activation) using costly and limited quantities of cell energy sources (e.g., ATP) in the soil ecosystem. IMPORTANCE The bacterial metabolic potential to synthesize diverse secondary metabolites in the environment has been revealed by recent (meta)genomics of both unculturable and culturable bacteria. These studies imply that bacteria are continuously exposed to harmful chemical compounds in the environment. Streptomyces spp. contain antibiotic efflux pumps and SM biosynthetic gene clusters. However, the mechanism by which soil bacteria, including Streptomyces, survive against toxic compounds in the environment remains unclear. Here, we identified three sets of TFR-ABC transporter genes in Streptomyces coelicolor A3(2). We found that each TFR controlled the expression of respective ABC transporter, and the expression of all ABC transporters negatively impacted SM production and increased antibiotic resistance. Notably, bioinformatic analysis indicated that these TFR-ABC transporter gene sets are highly conserved and widely distributed in the genome of Streptomyces species, indicating the importance of systematic regulation that directs antibiotic production and xenobiotic excretion.
    2023年05月, Applied and Environmental Microbiology, 89(3) (3), e0182222, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Takeshi Tsunoda, Shumpei Asamizu, Taifo Mahmud
    Acarbose, a pseudotetrasaccharide produced by several strains of Actinoplanes and Streptomyces, is an α-glucosidase inhibitor clinically used to control type II diabetes. Bioinformatic analysis of the biosynthetic gene clusters of acarbose in Actinoplanes sp. SE50/110 (the acb cluster) and Streptomyces glaucescens GLA.O (the gac cluster) revealed their distinct genetic organizations and presumably biosynthetic pathways. However, to date, only the acarbose pathway in the SE50/110 strain has been extensively studied. Here, we report that GacI, one of the proteins that appear to be different between the two pathways, is a bifunctional glycosyltransferase family 5 (GT5)-phosphatase (PP) enzyme that functions at two different steps in acarbose biosynthesis in S. glaucescens GLA.O. In the acb pathway, the GT and the PP reactions are performed by two different enzymes. Truncated GacI proteins having only the GT or the PP domain showed comparable catalytic activity with the full-length GacI, indicating that domain separation does not significantly affect their respective catalytic activity. GacI, which is widely distributed in many Streptomyces, represents the first example of naturally occurring GT5-PP bifunctional enzymes biochemically characterized.
    American Chemical Society (ACS), 2022年11月, Biochemistry, 61(22) (22), 2628 - 2635, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Shumpei Asamizu, Shinta Ijichi, Shotaro Hoshino, Hansaem Jo, Hidenori Takahashi, Yuko Itoh, Sohkichi Matsumoto, Hiroyasu Onaka
    Ribosomally synthesized and posttranslationally modified peptides (RiPPs) with polar-functionalized fatty acyl groups are a rarely found untapped class of natural products. Although polar-functionalized fatty-acylated RiPPs (PFARs) have potential as antimicrobial agents, the repertoire is still limited. Therefore, expanding the chemical space is expected to contribute to the development of pharmaceutical agents. In this study, we performed genome mining and stable isotope-guided comparative metabolomics to discover new PFAR natural products. We focused on the feature that PFARs incorporate l-arginine or l-lysine as the starter unit of the fatty acyl group and fed 13C6,15N4-l-arginine or 13C6,15N2-l-lysine to bacterial cultures. Metabolites were extracted and compared with those extracted from nonlabeled l-arginine or l-lysine fed cultures. We identified putative PFARs and successfully isolated solabiomycin A and B from Streptomyces lydicus NBRC 13 058 and albopeptin B from Streptomyces nigrescens HEK616, which contained a sulfoxide group in the labionin moiety. The gene disruption experiment indicated that solS, which encodes a putative flavin adenine dinucleotide (FAD)-nicotinamide adenine dinucleotide (phosphate) (NAD(P))-binding protein, is involved in the sulfoxidation of aryl sulfides. The solabiomycins showed antibacterial activity against Gram-positive bacteria, including Mycobacterium tuberculosis H37Rv with a minimum 95% inhibitory concentration (MIC95) of 3.125 μg/mL, suggesting their potential as antituberculosis agents.
    American Chemical Society (ACS), 2022年10月, ACS Chemical Biology, 17(10) (10), 2936 - 2944, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Shumpei Asamizu, Abrory Agus Cahya Pramana, Sung-Jin Kawai, Yoshichika Arakawa, Hiroyasu Onaka
    To investigate the potential for secondary metabolite biosynthesis by Streptomyces species, we employed a coculture method to discover natural bioactive products and identified specific antibacterial activity from a combined-culture of Streptomyces hygroscopicus HOK021 and Tsukamurella pulmonis TP-B0596. Molecular networking using ultrahigh performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS) data revealed a specific clade of metabolites in this combined-culture that were not detected in both monocultures. Using the chemical profiles, a previously unidentified conjugate between FabF inhibitor and catechol-type siderophore was successfully identified and named harundomycin A. Harundomycin A was a conjugate between the 2,4-dihydroxy-3-aminobenzoate moiety of platensimycin and N,N'-bis(2,3-dihydroxybenzoyl)-O-seryl-cysteine (bisDHBA-Ser-Cys) with a thioester linkage. Along with the production of harundomycin A, platensimycin, its thiocarboxylic acid form thioplatensimycin, enterobactin, and its degradation product N,N'-bis(2,3-dihydroxybenzoyl)-O-l-seryl-dehydroalanine (bisDHBA-Ser-Dha) were also induced in the combined-culture. Genomic data of S. hygroscopicus HOK021 and T. pulmonis TP-B0596 indicated that strain HOK021 possessed biosynthetic gene clusters for both platensimycin and enterobactin, and thereby revealed that T. pulmonis stimulates HOK021 and acts as an inducer of both of these metabolites. Although the harundomycin A was modified by bulky bisDHBA-Ser-Cys, responsible for the binding to the target molecule FabF, it showed a similar antibacterial spectrum to platensimycin, including against methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci, suggesting that the pharmacophore is platensimycin. Additionally, Chrome Azurol S assay showed that harundomycin A possesses ferric iron-chelating activity comparable to that of enterobactin. Our study demonstrated the transformation of existing natural products to bifunctional molecules driven by bacterial interaction.
    American Chemical Society (ACS), 2022年09月, ACS Chemical Biology, 17(9) (9), 2664 - 2672, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Masaomi Yanagisawa, Shumpei Asamizu, Katsuya Satoh, Yutaka Oono, Hiroyasu Onaka
    Streptomyces lividans TK23 interacts with mycolic acid-containing bacteria (MACB), such as Tsukamurella pulmonis TP-B0596, and this direct cell contact activates its secondary metabolism (e.g., the production of undecylprodigiosin: RED). Here, we employed carbon (12C5+) ion beam-induced mutagenesis to investigate the signature of induced point mutations and further identify the gene(s) responsible for the production of secondary metabolites induced by T. pulmonis. We irradiated spores of the Streptomyces coelicolor strain JCM4020 with carbon ions to generate a mutant library. We screened the RED production-deficient mutants of S. coelicolor by mixing them with T. pulmonis TP-B0596 on agar plates, identifying the red/white phenotype of the growing colonies. Through this process, we selected 59 RED-deficient mutants from around 152,000 tested spores. We resequenced the genomes of 16 mutants and identified 44 point mutations, which revealed the signatures induced by 12C5+-irradiation. Via gene complementation experiments, we also revealed that two genes-glutamate synthase (gltB) and elongation factor G (fusA)-are responsible for the reduced production of RED.
    2022年07月, PLoS One, 17(7) (7), e0270379, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Manami Kato, Shumpei Asamizu, Hiroyasu Onaka
    Abstract Co-culture is an efficient strategy for natural product discovery. We have used mycolic acid-containing bacteria (MACB) Tsukamurella pumonis TP-B0596 to induce secondary metabolism by actinomycetes and have found several natural products. We also observed that MACB attached to the mycelium of Streptomyces lividans forming coaggregates during combined-culture. This stimulated interest in the interactions among actinomycetes and MACB, and we found that soil isolated cultures contained a mixture of actinomycetes and MACB. Our previously observed interactions were the result of selective screening and combination of bacteria in the lab, which warranted investigation of the existence of these interactions in the natural soil environment. Therefore, in this paper, we report the interaction between a co-isolated natural pair of actinomycetes and MACB in terms of morphology and metabolic changes. A natural pair of actinomycetes and MACB co-aggregated in liquid culture and showed metabolic changes. Interestingly, co-aggregated actinomycetes and MACB were re-isolated from soil with no obvious morphological colony differences from the colony of a single strain. The results demonstrate that there is a stochastic chance of picking colonies containing co-aggregated actinomycetes and MACB, which suggests that the pair can exist in co-aggregate form in the soil environment and interact with each other.
    Springer Science and Business Media LLC, 2022年05月, Scientific Reports, 12(1) (1), 7222, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Ayumi Hikima, Shumpei Asamizu, Hiroyasu Onaka, Huiping Zhang, Hiroshi Tomoda, Nobuhiro Koyama
    During our screening for antibiotics against Mycobacterium avium complex (MAC) with a mass spectrometry network-based indexing approach, a new compound named kimidinomycin was isolated from the culture broth of Streptomyces sp. KKTA-0263 by solvent extraction, HP20 column chromatography, and preparative HPLC. From the structural elucidation, the compound possesses a 38-membered macrolide structure with an N-methylguanidyl group at the terminal side chain. The compound exhibited antimycobacterial activity against M. avium, M. intracellulare, M. smegmatis, and M. bovis BCG with respective MIC values of 12.5, 0.78, 12.5, and 25.0 µg ml-1.
    2022年02月, The Journal of Antibiotics, 75(2) (2), 72 - 76, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Yoshiki Asai, Tomoshige Hiratsuka, Miyu Ueda, Yumi Kawamura, Shumpei Asamizu, Hiroyasu Onaka, Manabu Arioka, Shinichi Nishimura, Minoru Yoshida
    Ferrichromes are a family of fungal siderophores with cyclic hexapeptide structures. Most fungi produce one or two ferrichrome-type siderophores. Acremonium persicinum MF-347833 produces ferrichrome-like potent Trojan horse antifungal antibiotics ASP2397 and AS2488053, the aluminum- and iron-chelating forms of AS2488059, respectively. Here, we show by gene sequencing followed by gene deletion experiments that A. persicinum MF-347833 possesses two nonribosomal peptide synthetase genes responsible for AS2488059 and ferricrocin assembly. AS2488059 was produced under iron starvation conditions and excreted into the media to serve as a defense metabolite and probably an iron courier. In contrast, ferricrocin was produced under iron-replete conditions and retained inside the cells, likely serving as an iron-sequestering molecule. Notably, the phylogenetic analyses suggest the different evolutionary origin of AS2488059 from that of conventional ferrichrome-type siderophores. Harnessing two ferrichrome-type siderophores with distinct biological properties may give A. persicinum a competitive advantage for surviving the natural environment.
    2022年01月, ACS Chemical Biology, 17(1) (1), 207 - 216, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Toshiki Nagakubo, Tatsuya Yamamoto, Shumpei Asamizu, Masanori Toyofuku, Nobuhiko Nomura, Hiroyasu Onaka
    Extracellular contractile injection systems (eCISs) are structurally similar to headless phages and are versatile nanomachines conserved among diverse classes of bacteria. Herein, Streptomyces species, which comprise filamentous Gram-positive bacteria and are ubiquitous in soil, were shown to produce Streptomyces phage tail-like particles (SLPs) from eCIS-related genes that are widely conserved among Streptomyces species. In some Streptomyces species, these eCIS-related genes are regulated by a key regulatory gene, which is essential for Streptomyces life cycle and is involved in morphological differentiation and antibiotic production. Deletion mutants of S. lividans of the eCIS-related genes appeared phenotypically normal in terms of morphological differentiation and antibiotic production, suggesting that SLPs are involved in other aspects of Streptomyces life cycle. Using co-culture method, we found that colonies of SLP-deficient mutants of S. lividans were more severely invaded by fungi, including Saccharomyces cerevisiae and Schizosaccharomyces pombe. In addition, microscopic and transcriptional analyses demonstrated that SLP expression was elevated upon co-culture with the fungi. In contrast, co-culture with Bacillus subtilis markedly decreased SLP expression and increased antibiotic production. Our findings demonstrate that in Streptomyces, eCIS-related genes affect microbial competition, and the patterns of SLP expression can differ depending on the competitor species.
    2021年10月, Scientific Reports, 11(1) (1), 20116, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Ryosuke Kozakai, Takuto Ono, Shotaro Hoshino, Hidenori Takahashi, Yohei Katsuyama, Yoshinori Sugai, Taro Ozaki, Kazuya Teramoto, Kanae Teramoto, Koichi Tanaka, Ikuro Abe, Shumpei Asamizu, Hiroyasu Onaka
    Fusions of fatty acids and peptides expand the structural diversity of natural products; however, polyketide/ribosomally synthesized and post-translationally modified peptides (PK/RiPPs) hybrid lipopeptides are relatively rare. Here we report a family of PK/RiPPs called goadvionins, which inhibit the growth of Gram-positive bacteria, and an acyltransferase, GdvG, which catalyses the condensation of the PK and RiPP moieties. Goadvionin comprises a trimethylammonio 32-carbon acyl chain and an eight-residue RiPP with an avionin structure. The positions of six hydroxyl groups and one double bond in the very-long acyl chain were determined by radical-induced dissociation tandem mass spectrometry, which collides radical ion species to generate C-C bond cleavage fragments. GdvG belongs to the Gcn5-related N-acetyltransferase superfamily. Unlike conventional acyltransferases, GdvG transfers a very long acyl chain that is tethered to an acyl carrier protein to the N-terminal amino group of the RiPP moiety. gdvG homologues flanked by PK/fatty acid and RiPP biosynthesis genes are widely distributed in microbial species, suggesting that acyltransferase-catalysed condensation of PKs and RiPPs is a general strategy in biosynthesis of similar lipopeptides.
    2020年09月, Nature chemistry, 12(9) (9), 869 - 877, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Alexander A Vinogradov, Morito Shimomura, Yuki Goto, Taro Ozaki, Shumpei Asamizu, Yoshinori Sugai, Hiroaki Suga, Hiroyasu Onaka
    Lactazole A is a cryptic thiopeptide from Streptomyces lactacystinaeus, encoded by a compact 9.8 kb biosynthetic gene cluster. Here, we establish a platform for in vitro biosynthesis of lactazole A, referred to as the FIT-Laz system, via a combination of the flexible in vitro translation (FIT) system with recombinantly produced lactazole biosynthetic enzymes. Systematic dissection of lactazole biosynthesis reveals remarkable substrate tolerance of the biosynthetic enzymes and leads to the development of the minimal lactazole scaffold, a construct requiring only 6 post-translational modifications for macrocyclization. Efficient assembly of such minimal thiopeptides with FIT-Laz opens access to diverse lactazole analogs with 10 consecutive mutations, 14- to 62-membered macrocycles, and 18 amino acid-long tail regions, as well as to hybrid thiopeptides containing non-proteinogenic amino acids. This work suggests that the minimal lactazole scaffold is amenable to extensive bioengineering and opens possibilities to explore untapped chemical space of thiopeptides.
    2020年05月, Nature communications, 11(1) (1), 2272, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Lu-Min Vaario, Shumpei Asamizu, Tytti Sarjala, Norihisa Matsushita, Hiroyasu Onaka, Yan Xia, Hiroyuki Kurokochi, Shin-Ichi Morinaga, Jian Huang, Shijie Zhang, Chunlan Lian
    Tricholoma matsutake is known to be the dominant fungal species in matsutake fruitbody neighboring (shiro) soil. To understand the mechanisms behind matsutake dominance, we studied the bacterial communities in matsutake dominant shiro soil and non-shiro soil, isolated the strains of Streptomyces from matsutake mycorrhizal root tips both from shiro soil and from the Pinus densiflora seedlings cultivated in shiro soil. Further, we investigated three Streptomyces spp. for their ability to inhibit fungal growth and Pinus densiflora seedling root elongation as well as two strains for their antifungal and antioxidative properties. Our results showed that Actinobacteria was the most abundant phylum in shiro soil. However, the differences in the Actinobacterial community composition (phylum or order level) between shiro and non-shiro soils were not significant, as indicated by PERMANOVA analyses. A genus belonging to Actinobacteria, Streptomyces, was present on the matsutake mycorrhizas, although in minority. The two antifungal assays revealed that the broths of three Streptomyces spp. had either inhibitory, neutral or promoting effects on the growth of different forest soil fungi as well as on the root elongation of the seedlings. The extracts of two strains, including one isolated from the P. densiflora seedlings, inhibited the growth of either pathogenic or ectomycorrhizal fungi. The effect depended on the medium used to cultivate the strains, but not the solvent used for the extraction. Two Streptomyces spp. showed antioxidant activity in one out of three assays used, in a ferric reducing antioxidant power assay. The observed properties seem to have several functions in matsutake shiro soil and they may contribute to the protection of the shiro area for T. matsutake dominance.
    SPRINGER, 2020年04月, Symbiosis, 81, 1 - 13, 英語
    [査読有り]
    研究論文（学術雑誌）

  • Ryosuke Sugiyama, Takahiro Nakatani, Shinichi Nishimura, Kei Takenaka, Taro Ozaki, Shumpei Asamizu, Hiroyasu Onaka, Hideaki Kakeya
    Organisms often produce secondary metabolites as a mixture of biosynthetically related congeners. However, why are metabolites with minor chemical variations produced simultaneously? 5-Alkyl-1,2,3,4-tetrahydroquinolines (5aTHQs) are small, lipophilic metabolites produced by Streptomyces nigrescens HEK616 when cultured with Tsukamurella pulmonis TP-B0596. A mixture of 5aTHQs forms aggregates that show enhanced membrane affinity and biological activity. The ability to form aggregates and membrane-binding activity is regulated by the length of the alkyl chains. Aggregates with long alkyl chains were too stable to fuse with lipid membranes. However, if inactive 5aTHQ congener was mixed with active congener, the mixture showed increased membrane affinity, enabling cellular entry and biological activity. Therefore, it is shown that sloppiness in a biosynthetic pathway, by which minor structural variations can be produced, is functionally rational, as the metabolites show synergistic action.
    2019年09月, Angewandte Chemie (International ed. in English), 58(38) (38), 13486 - 13491, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Taro Ozaki, Ryosuke Sugiyama, Morito Shimomura, Shinichi Nishimura, Shumpei Asamizu, Yohei Katsuyama, Hideaki Kakeya, Hiroyasu Onaka
    5-Alkyl-1,2,3,4-tetrahydroquinolines (5aTHQs) and streptoaminals (STAMs) are natural products isolated from the combined-culture of Streptomyces nigrescens HEK616 and Tsukamurella pulmonis TP-B0596. Despite their unique structures, their biosynthetic pathway has yet to be elucidated. In the present study, we conducted a feeding experiment using 13C-labeled acetates and demonstrated that 5aTHQs are likely synthesized by the action of polyketide synthase (PKS). Based on this observation, we identified the biosynthetic gene cluster for 5aTHQs. Interestingly, the same gene cluster was also responsible for the structurally-distinct STAMs. The gene cluster contains nine genes encoding one acyl carrier protein, two sets of ketosynthases (KSs) and chain length factors (CLFs), one aminotransferase/reductase bifunctional protein, two ketoreductases, and one thioesterase. KSs and CLFs are classified into the phylogenetically distinct clades from those of known type II PKSs. Heterologous expression of the biosynthetic genes and subsequent gene inactivation clearly indicated that all of the nine genes were required for the biosynthesis of both compounds. In the proposed biosynthetic pathway, chain elongation by PKS, reductive cleavage of a thioester bond, and subsequent transamination generate the core skeleton of both compounds. Differences in the oxidation states of the products result in a distinct cyclization mode to yield 5aTHQs and STAMs.
    2019年02月, Organic & Biomolecular Chemistry, 17(9) (9), 2370 - 2378, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • 尾仲 宏康, 丸山 潤一, 浅水 俊平, 黒岩 真弓, 北本 勝ひこ, 山田 雅人, 五島 徹也, 赤尾 健
    富山県高岡市産大麦麦芽より醸造に適した「とやま産まれの酵母」を分離した。本菌株はITS領域解析の結果，Saccharomyces cerevsiae NBRC2114（Kotobukiya whisky No. 1）に最も近い配列を有することが明らかとなった。清酒小仕込み試験による醸造特性解析の結果，とやま産まれの酵母のもろみ日数はK701よりも3日長くなったものの，両者の到達アルコール濃度はほぼ同じ18%であった。K701と比較し，有機酸組成では酢酸，リンゴ酸，コハク酸の含有量が顕著に異なっており，揮発性物質では高級アルコール含量が高く，エステル類の含量が低かった。アミノ酸度はとやま酵母の方が顕著に高かった。これらの性質が官能特性評価にも現れており，K701に比較して，酸味の違いや，旨み，重い，雑味などの味の多さに特徴がある個性的な酒質であった。
    とやま産まれの酵母による実地醸造は成政酒造にて，これまでに5回行われており，試行錯誤の末，現在では比較的濃醇でキレのある酒質となっている。また，本酵母は野生酵母で特定の酒類醸造に特化した育種もなされていないため，富山県地域のクラフトビール，ワイン，ウイスキーの醸造にも利用されるに至っている。
    公益財団法人 日本醸造協会, 2019年, 日本醸造協会誌, 114(10) (10), 645 - 653, 日本語

  • Shotaro Hoshino, Chin Piow Wong, Masahiro Ozeki, Huiping Zhang, Fumiaki Hayashi, Takayoshi Awakawa, Shumpei Asamizu, Hiroyasu Onaka, Ikuro Abe
    New polycyclic tetramate macrolactams, Umezawamides A (1) and B (2) were isolated from a combined-culture of Umezawaea sp. RD066910 and mycolic-acid containing bacterium Tsukamurella pulmonis TP-B0596. Their planar structures and partial stereochemistries were determined based on the spectroscopic analysis, MMFF conformational search, and ECD calculations. Umezawamides are the first secondary metabolites isolated from the genus Umezawaea and they exhibited cytotoxicities to P388 murine leukemia cells. Furthermore, umezawamide A (1) showed growth inhibitory activity against Candida albicans.
    2018年07月, The Journal of Antibiotics, 71(7) (7), 653 - 657, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Shotaro Hoshino, Masahiro Okada, Takayoshi Awakawa, Shumpei Asamizu, Hiroyasu Onaka, Ikuro Abe
    Two novel macrolactams, dracolactams A and B, were identified from a combined-culture of Micromonospora species and a mycolic-acid containing bacterium (MACB). Their structures and stereochemistries were completely assigned, based on spectroscopic analyses and chemical derivatization. Both dracolactams were probably generated from a common macrolactam precursor produced by the Micromonospora species. In this combined-culture system, MACB is likely to activate cryptic oxidase genes in the Micromonospora species and induce the downstream polyene macrolactam cyclization.
    2017年09月, Organic letters, 19(18) (18), 4992 - 4995, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Andrew R Osborn, Kelsey M Kean, Khaled M Alseud, Khaled H Almabruk, Shumpei Asamizu, Janet A Lee, P Andrew Karplus, Taifo Mahmud
    2-Epi-5-epi-valiolone synthase (EEVS), a C7-sugar phosphate cyclase (SPC) homologous to 3-dehydroquinate synthase (DHQS), was discovered during studies of the biosynthesis of the C7N-aminocyclitol family of natural products. EEVS was originally thought to be present only in certain actinomycetes, but analyses of genome sequences showed that it is broadly distributed in both prokaryotes and eukaryotes, including vertebrates. Another SPC, desmethyl-4-deoxygadusol synthase (DDGS), was later discovered as being involved in the biosynthesis of mycosporine-like amino acid sunscreen compounds. Current database annotations are quite unreliable, with many EEVSs reported as DHQS, and most DDGSs reported as EEVS, DHQS, or simply hypothetical proteins. Here, we identify sequence features useful for distinguishing these enzymes, report a crystal structure of a representative DDGS showing the high similarity of the EEVS and DDGS enzymes, identify notable active site differences, and demonstrate the importance of two of these active site residues for catalysis by point mutations. Further, we functionally characterized two representatives of a distinct clade equidistant from known EEVS and known DDGS groups and show them to be authentic EEVSs. Moreover, we document and discuss the distribution of genes that encode EEVS and DDGS in various prokaryotes and eukaryotes, including pathogenic bacteria, plant symbionts, nitrogen-fixing bacteria, myxobacteria, cyanobacteria, fungi, stramenopiles, and animals, suggesting their broad potential biological roles in nature.
    2017年04月, ACS Chemical Biology, 12(4) (4), 979 - 988, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Taro Ozaki, Kona Yamashita, Yuki Goto, Morito Shimomura, Shohei Hayashi, Shumpei Asamizu, Yoshinori Sugai, Haruo Ikeda, Hiroaki Suga, Hiroyasu Onaka
    Goadsporin (GS) is a member of ribosomally synthesized and post-translationally modified peptides (RiPPs), containing an N-terminal acetyl moiety, six azoles and two dehydroalanines in the peptidic main chain. Although the enzymes involved in GS biosynthesis have been defined, the principle of how the respective enzymes control the specific modifications remains elusive. Here we report a one-pot synthesis of GS using the enzymes reconstituted in the 'flexible' in vitro translation system, referred to as the FIT-GS system. This system allows us to readily prepare not only the precursor peptide from its synthetic DNA template but also 52 mutants, enabling us to dissect the modification determinants of GodA for each enzyme. The in vitro knowledge has also led us to successfully produce designer GS analogues in vivo. The methodology demonstrated in this work is also applicable to other RiPP biosynthesis, allowing us to rapidly investigate the principle of modification events with great ease.
    NATURE PUBLISHING GROUP, 2017年02月, Nature communications, 8, 14207, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Ryosuke Sugiyama, Shinichi Nishimura, Taro Ozaki, Shumpei Asamizu, Hiroyasu Onaka, Hideaki Kakeya
    A series of lipidic spirohemiaminals, designated streptoaminals, is reported. These were discovered by surveying the unique molecular signatures identified in the mass spectrometry data of the combined-culture broth of Streptomyces nigrescens HEK616 and Tsukamurella pulmonis TP-B0596. Mass spectrometry analysis showed that streptoaminals appeared as a cluster of ion peaks, which were separated by 14 mass unit intervals, implying the presence of alkyl chains of different lengths. The chemical structures of these compounds were elucidated by spectroscopic analysis and total synthesis. Streptoaminals with globular structures showed broad antimicrobial activities, whereas the planar structures of the 5-alkyl-1,2,3,4-tetrahydroquinolines found in the same combined-culture did not. This work shows the application of microbes as reservoirs for a range of chemical scaffolds.
    2016年08月, Angewandte Chemie (International ed. in English), 55(35) (35), 10278 - 10282, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Taro Ozaki, Yukari Kurokawa, Shohei Hayashi, Naoya Oku, Shumpei Asamizu, Yasuhiro Igarashi, Hiroyasu Onaka
    Dehydroalanines in goadsporin are proposed to be formed by GodF and GodG, which show slight homology to the N-terminal glutamylation and C-terminal elimination domains, respectively, of LanB, a class I lanthipeptide dehydratase. Although similar, separated-type LanBs are conserved among thiopeptides and indispensable for their biosynthesis and biological activities, these enzymes had not yet been characterized. Here, we identified goadsporin B, which has unmodified Ser4 and Ser14, from both godF and godG disruptants. The godG disruptant also produced goadsporin C, a glutamylated-Ser4 variant of goadsporin B. These results suggested that dehydroalanines are formed by glutamylation and glutamate elimination. NMR analysis revealed for the first time that the glutamyl group was attached to a serine via an ester bond, by the catalysis of LanB-type enzymes. Our findings provide insights into the function of separated-type LanBs involved in the biosynthesis of goadsporin and thiopeptides.
    2016年02月, Chembiochem : a European journal of chemical biology, 17(3) (3), 218 - 223, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Shumpei Asamizu, Taro Ozaki, Kanae Teramoto, Katsuya Satoh, Hiroyasu Onaka
    Co-culture of Streptomyces with mycolic acid-containing bacteria (MACB), which we termed "combined-culture," alters the secondary metabolism pattern in Streptomyces and has been a useful method for the discovery of bioactive natural products. In the course of our investigation to identify the inducing factor(s) of MACB, we previously observed that production of pigments in Streptomyces lividans was not induced by factors such as culture extracts or mycolic acids. Although dynamic changes occurred in culture conditions because of MACB, the activation of pigment production by S. lividans was observed in a limited area where both colonies were in direct contact. This suggested that direct attachment of cells is a requirement and that components on the MACB cell membrane may play an important role in the response by S. lividans. Here we examined whether this response was influenced by dead MACB that possess intact mycolic acids assembled on the outer cell membrane. Formaldehyde fixation and γ-irradiation were used to prepare dead cells that retain their shape and mycolic acids of three MACB species: Tsukamurella pulmonis, Rhodococcus erythropolis, and Rhodococcus opacus. Culturing tests verified that S. lividans does not respond to the intact dead cells of three MACB. Observation of combined-culture by scanning electron microscopy (SEM) indicated that adhesion of live MACB to S. lividans mycelia were a significant interaction that resulted in formation of co-aggregation. In contrast, in the SEM analysis, dead cells were not observed to adhere. Therefore, direct attachment by live MACB cells is proposed as one of the possible factors that causes Streptomyces to alter its specialized metabolism in combined-culture.
    2015年11月, PLoS One, 10(11) (11), e0142372, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Hiroyasu Onaka, Taro Ozaki, Yukiko Mori, Masumi Izawa, Shohei Hayashi, Shumpei Asamizu
    2015年09月, The Journal of Antibiotics, 68(9) (9), 594 - 597, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Andrew R Osborn, Khaled H Almabruk, Garrett Holzwarth, Shumpei Asamizu, Jane LaDu, Kelsey M Kean, P Andrew Karplus, Robert L Tanguay, Alan T Bakalinsky, Taifo Mahmud
    Ultraviolet-protective compounds, such as mycosporine-like amino acids (MAAs) and related gadusols produced by some bacteria, fungi, algae, and marine invertebrates, are critical for the survival of reef-building corals and other marine organisms exposed to high-solar irradiance. These compounds have also been found in marine fish, where their accumulation is thought to be of dietary or symbiont origin. In this study, we report the unexpected discovery that fish can synthesize gadusol de novo and that the analogous pathways are also present in amphibians, reptiles, and birds. Furthermore, we demonstrate that engineered yeast containing the fish genes can produce and secrete gadusol. The discovery of the gadusol pathway in vertebrates provides a platform for understanding its role in these animals, and the possibility of engineering yeast to efficiently produce a natural sunscreen and antioxidant presents an avenue for its large-scale production for possible use in pharmaceuticals and cosmetics.
    2015年05月, eLife, 4, e05919, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Ryosuke Sugiyama, Shinichi Nishimura, Taro Ozaki, Shumpei Asamizu, Hiroyasu Onaka, Hideaki Kakeya
    Eight novel 5-alkyl-1,2,3,4-tetrahydroquinolines (5aTHQs) bearing different side chains have been isolated from a combined culture of Streptomyces nigrescens HEK616 and Tsukamurella pulmonis TP-B0596. The chemical structures including the absolute configuration were elucidated by spectroscopic analysis and total synthesis. 5aTHQs inhibited the growth of wild-type fission yeast while only weakly inhibiting the growth of several mutant strains synthesizing premature ergosterol. These results demonstrate that 5aTHQs are novel antifungals that may target cell membranes.
    2015年04月, Organic letters, 17(8) (8), 1918 - 1921, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Kelsey M Kean, Sara J Codding, Shumpei Asamizu, Taifo Mahmud, P Andrew Karplus
    Sedoheptulose 7-phosphate cyclases (SH7PCs) encompass three enzymes involved in producing the core cyclitol structures of pseudoglycosides and similar bioactive natural products. One such enzyme is ValA from Streptomyces hygroscopicus subsp. jinggangensis 5008, which makes 2-epi-5-epi-valiolone as part of the biosynthesis of the agricultural antifungal agent validamycin A. We present, as the first SH7PC structure, the 2.1 Å resolution crystal structure of ValA in complex with NAD+ and Zn2+ cofactors. ValA has a fold and active site organization resembling those of the sugar phosphate cyclase dehydroquinate synthase (DHQS) and contains two notable, previously unrecognized interactions between NAD+ and Asp side chains conserved in all sugar phosphate cyclases that may influence catalysis. Because the domains of ValA adopt a nearly closed conformation even though no sugar substrate is present, comparisons with a ligand-bound DHQS provide a model for aspects of substrate binding. One striking active site difference is a loop that adopts a distinct conformation as a result of an Asp→Asn change with respect to DHQS and alters the identity and orientation of a key Arg residue. This and other active site differences in ValA are mostly localized to areas where the ValA substrate differs from that of DHQS. Sequence comparisons with a second SH7PC making a product with distinct stereochemistry lead us to postulate that the product stereochemistry of a given SH7PC is not the result of events taking place during catalysis but is accomplished by selective binding of either the α or β pyranose anomer of the substrate.
    2014年07月, Biochemistry, 53(26) (26), 4250 - 4260, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Shohei Hayashi, Taro Ozaki, Shumpei Asamizu, Haruo Ikeda, Satoshi Ōmura, Naoya Oku, Yasuhiro Igarashi, Hiroshi Tomoda, Hiroyasu Onaka
    Although >100 thiopeptides have been discovered, the number of validated gene clusters involved in their biosynthesis is lagging. We use genome mining to identify a silent thiopeptide biosynthetic gene cluster responsible for biosynthesis of lactazoles. Lactazoles are structurally unique thiopeptides with a 32-membered macrocycle and a 2-oxazolyl-6-thiazolyl pyridine core. We demonstrate that lactazoles originate from the simplest cluster, containing only six unidirectional genes (lazA to lazF). We show that lazC is involved in the macrocyclization process, leading to central pyridine moiety formation. Substitution of the endogenous promoter with a strong promoter results in an approximately 30-fold increase in lactazole A production and mutagenesis of lazC precursor gene in production of two analogs. Lactazoles do not exhibit antimicrobial activity but may modulate signaling cascades triggered by bone morphogenetic protein. Our approach facilitates the production of a more diverse set of thiopeptide structures, increasing the semisynthetic repertoire for use in drug development.
    2014年05月, Chemistry & Biology, 21(5) (5), 679 - 688, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Kentaro Haginaka, Shumpei Asamizu, Taro Ozaki, Yasuhiro Igarashi, Tamotsu Furumai, Hiroyasu Onaka
    Improving the productivity of secondary metabolites is highly beneficial for the utilization of natural products. Here, we found that gene duplication of the goadsporin biosynthetic gene locus resulted in hyper-production. Goadsporin is a linear azole containing peptide that is biosynthesized via a ribosome-mediated pathway in Streptomyces sp. TP-A0584. Recombinant strains containing duplicated or triplicated goadsporin biosynthetic gene clusters produced 1.46- and 2.25-fold more goadsporin than the wild-type strain. In a surrogate host, Streptomyces lividans, chromosomal integration of one or two copies of the gene cluster led to 342.7 and 593.5 mg/L of goadsporin production. Expression of godI, a self-resistance gene, and of godR, a pathway-specific transcriptional regulator, under a constitutive promoter gave 0.79- and 2.12-fold higher goadsporin production than the wild-type strain. Our experiments indicated that a proportional relationship exists between goadsporin production per culture volume and the copy number of the biosynthetic gene cluster.
    2014年03月, Bioscience, Biotechnology, and Biochemistry, 78(3) (3), 394 - 399, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Shumpei Asamizu, Mostafa Abugreen, Taifo Mahmud
    Through a combination of genome mining, gene knockouts, and comparative metabolomic studies, a new biosynthetic pathway to validoxylamine A has been discovered in Actinosynnema mirum DSM 43827. The study revealed the rich diversity of biosynthetic pathways to pseudosugar-containing natural products. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
    2013年09月, Chembiochem : a European journal of chemical biology, 14(13) (13), 1548 - 1551, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Shumpei Asamizu, Satoshi Hirano, Hiroyasu Onaka, Hiroyuki Koshino, Yoshitsugu Shiro, Shingo Nagano
    2012年11月, Chembiochem : a European journal of chemical biology, 13(17) (17), 2495 - 2500, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Khaled H Almabruk, Shumpei Asamizu, Ada Chang, Sheril G Varghese, Taifo Mahmud
    From A to B: Through detailed biochemical investigations, we discovered that VldW, an α-ketoglutarate/Fe(II)-dependent dioxygenase, regioselectively hydroxylates validamycin A to validamycin B. The results provide insights into the biosynthesis of hydroxylated validamycins and could be used to control the metabolic outcomes of the validamycin pathway.
    2012年10月, Chembiochem : a European journal of chemical biology, 13(15) (15), 2209 - 2211, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Michael C Cavalier, Young-Sun Yim, Shumpei Asamizu, David Neau, Khaled H Almabruk, Taifo Mahmud, Yong-Hwan Lee
    The pseudo-glycosyltransferase VldE catalyzes non-glycosidic C-N coupling between an unsaturated cyclitol and a saturated aminocyclitol with the conservation of the stereochemical configuration of the substrates to form validoxylamine A 7'-phosphate, the biosynthetic precursor of the antibiotic validamycin A. To study the molecular basis of its mechanism, the three-dimensional structures of VldE from Streptomyces hygroscopicus subsp. limoneus was determined in apo form, in complex with GDP, in complex with GDP and validoxylamine A 7'-phosphate, and in complex with GDP and trehalose. The structure of VldE with the catalytic site in both an "open" and "closed" conformation is also described. With these structures, the preferred binding of the guanine moiety by VldE, rather than the uracil moiety as seen in OtsA could be explained. The elucidation of the VldE structure in complex with the entirety of its products provides insight into the internal return mechanism by which catalysis occurs with a net retention of the stereochemical configuration of the donated cyclitol.
    2012年09月, PLoS One, 7(9) (9), e44934, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Shumpei Asamizu, Pengfei Xie, Corey J Brumsted, Patricia M Fla, Taifo Mahmud
    Sedoheptulose 7-phosphate cyclases are enzymes that utilize the pentose phosphate pathway intermediate, sedoheptulose 7-phosphate, to generate cyclic precursors of many bioactive natural products, such as the antidiabetic drug acarbose, the crop protectant validamycin, and the natural sunscreens mycosporine-like amino acids. These proteins are phylogenetically related to the dehydroquinate (DHQ) synthases from the shikimate pathway and are part of the more recently recognized superfamily of sugar phosphate cyclases, which includes DHQ synthases, aminoDHQ synthases, and 2-deoxy-scyllo-inosose synthases. Through genome mining and biochemical studies, we identified yet another subset of DHQS-like proteins in the actinomycete Actinosynnema mirum and the myxobacterium Stigmatella aurantiaca DW4/3-1. These enzymes catalyze the conversion of sedoheptulose 7-phosphate to 2-epi-valiolone, which is predicted to be an alternative precursor for aminocyclitol biosynthesis. Comparative bioinformatics and biochemical analyses of these proteins with 2-epi-5-epi-valiolone synthases (EEVS) and desmethyl-4-deoxygadusol synthases (DDGS) provided further insights into their genetic diversity, conserved amino acid sequences, and plausible catalytic mechanisms. The results further highlight the uniquely diverse DHQS-like sugar phosphate cyclases, which may provide new tools for chemoenzymatic, stereospecific synthesis of various cyclic molecules.
    2012年07月, Journal of the American Chemical Society, 134(29) (29), 12219 - 12229, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Shumpei Asamizu, Yoshitsugu Shiro, Yasuhiro Igarashi, Shingo Nagano, Hiroyasu Onaka
    The diversity of indolocarbazole natural products results from the differences in oxidation states of the pyrroline ring moiety. In the biosynthetic pathways for staurosporine and rebeccamycin, two homologous enzymes having 64% identity, StaC and RebC, are responsible for the selective production of K252c, which has one oxo group at the pyrroline ring, and arcyriaflavin A, which has two. Although StaC has a FAD-binding motif, most StaC molecules do not contain FAD, and the protein cannot be reconstituted with FAD in vitro. In this study, we mutated Ala-118 in StaC by replacing a glutamine that is conserved in FAD monooxygenases, resulting in increased FAD content as well as catalytic activity. In addition, mutations around the substrate-binding sites of StaC and RebC can change the product selectivity. Specifically, StaC-N244R-V246T and RebC-F216V-R239N mutants produced substantial amounts of arcyriaflavin A and K252c, respectively.
    2011年11月, Bioscience, Biotechnology, and Biochemistry, 75(11) (11), 2184 - 2193, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Shumpei Asamizu, Jongtae Yang, Khaled H Almabruk, Taifo Mahmud
    Glycosyltransferases are ubiquitous in nature. They catalyze a glycosidic bond formation between sugar donors and sugar or nonsugar acceptors to produce oligo/polysaccharides, glycoproteins, glycolipids, glycosylated natural products, and other sugar-containing entities. However, a trehalose 6-phosphate synthase-like protein has been found to catalyze an unprecedented nonglycosidic C-N bond formation in the biosynthesis of the aminocyclitol antibiotic validamycin A. This dedicated 'pseudoglycosyltransferase' catalyzes a condensation between GDP-valienol and validamine 7-phosphate to give validoxylamine A 7'-phosphate with net retention of the 'anomeric' configuration of the donor cyclitol in the product. The enzyme operates in sequence with a phosphatase, which dephosphorylates validoxylamine A 7'-phosphate to validoxylamine A.
    2011年08月, Journal of the American Chemical Society, 133(31) (31), 12124 - 12135, 英語, 国際誌
    [査読有り]
    研究論文（学術雑誌）

  • Yong Wang, Hui Chen, Masatomo Makino, Yoshitsugu Shiro, Shingo Nagano, Shumpei Asamizu, Hiroyasu Onaka, Sason Shaik
    American Chemical Society (ACS), 2009年05月, Journal of the American Chemical Society, 131(19) (19), 6748 - 6762, 英語
    [査読有り]
    研究論文（学術雑誌）

  • Satoshi Hirano, Shumpei Asamizu, Hiroyasu Onaka, Yoshitsugu Shiro, Shingo Nagano
    Elsevier BV, 2008年03月, Journal of Biological Chemistry, 283(10) (10), 6459 - 6466, 英語
    [査読有り]
    研究論文（学術雑誌）

  • Masatomo Makino, Hiroshi Sugimoto, Yoshitsugu Shiro, Shumpei Asamizu, Hiroyasu Onaka, Shingo Nagano
    Staurosporine isolated from Streptomyces sp. TP-A0274 is a member of the family of indolocarbazole alkaloids that exhibit strong antitumor activity. A key step in staurosporine biosynthesis is the formation of the indolocarbazole core by intramolecular C–C bond formation and oxidative decarboxylation of chromopyrrolic acid (CPA) catalyzed by cytochrome P450 StaP (StaP, CYP245A1). In this study, we report x-ray crystal structures of CPA-bound and -free forms of StaP. Upon substrate binding, StaP adopts a more ordered conformation, and conformational rearrangements of residues in the active site are also observed. Hydrogen-bonding interactions of two carboxyl groups and T-shaped π–π interactions with indole rings hold the substrate in the substrate-binding cavity with a conformation perpendicular to the heme plane. Based on the crystal structure of StaP–CPA complex, we propose that C–C bond formation occurs through an indole cation radical intermediate that is equivalent to cytochrome c peroxidase compound I [Sivaraja M, Goodin DB, Smith M, Hoffman BM (1989) Science 245:738–740]. The subsequent oxidative decarboxylation reaction is also discussed based on the crystal structure. Our crystallographic study shows the first crystal structures of enzymes involved in formation of the indolocarbazole core and provides valuable insights into the process of staurosporine biosynthesis, combinatorial biosynthesis of indolocarbazoles, and the diversity of cytochrome P450 chemistry.
    Proceedings of the National Academy of Sciences, 2007年07月, Proceedings of the National Academy of Sciences, 104(28) (28), 11591 - 11596, 英語
    [査読有り]
    研究論文（学術雑誌）

  • Shumpei Asamizu, Yasuo Kato, Yasuhiro Igarashi, Hiroyasu Onaka
    Elsevier BV, 2007年04月, Tetrahedron Letters, 48(16) (16), 2923 - 2926, 英語
    [査読有り]
    研究論文（学術雑誌）

  • Shumpei Asamizu, Yasuo Kato, Yasuhiro Igarashi, Tamotsu Furumai, Hiroyasu Onaka
    Elsevier BV, 2006年01月, Tetrahedron Letters, 47(4) (4), 473 - 475, 英語
    [査読有り]
    研究論文（学術雑誌）

  • Hiroyasu ONAKA, Shumpei ASAMIZU, Yasuhiro IGARASHI, Ryuji YOSHIDA, Tamotsu FURUMAI
    Oxford University Press (OUP), 2005年01月, Bioscience, Biotechnology, and Biochemistry, 69(9) (9), 1753 - 1759, 英語
    [査読有り]
    研究論文（学術雑誌）

■ MISC
  • Manami Kato, Shumpei Asamizu, Hiroyasu Onaka
    Abstract Chemical cross talks betweenMycolicibacterium septicumHEK138M andBacillus subtilis168 affect the bacterial morphology ofStreptomyces variegatusHEK138A. We found thatS. variegatusexhibits unusual hyphae branching by the bacterial interaction. We aimed to elucidate the mechanism by performing activity guided purification of substances that induce the unusual cell morphology. We found that pyrogallol, a redox active aromatic small molecule induced significant hyphae branching inS. variegatusand the activity was also observed in some of otherStreptomycesspecies. Interestingly, the pyrogallol activity was diminished by adding catalase, which broke down H2O2. To further confirm the involvement, H2O2was tested and similar activity which induced hyphal branching was observed. This indicates that reactive oxygen species (ROS) generated by redox-active compound (RAC) is the inducing factor of hyphae branching. Further investigation revealed that pyrogallol was generated by NahG enzyme homolog ofM. septicumusing 2,3-dihydroxybenzoic acid as substrate by heterologous expression inE. coli. Moreover, co-culture with gene knock-out mutants revealed that 2,3-dihydroxybenzoic acid was supplied byB. subtilisproduced as intermediate of bacterial siderophore bacillibactin. Since the hyphae branching of vegetative mycelium can increase the density of filamentous network and consequently help secure the milieu in soil, our results suggested that those filamentous soil bacteria use ROS which can be supplied from plant derived RAC as a signal. As those RAC ubiquitously exist in soil environment, the system will be beneficial for sensing the nutrient sources in addition to the generally considered defensive response to oxidative stress. Importance The characterization of interactions between three or more bacteria are lacking as these interactions are visually imperceptible in general. Our current study revealed changes of morphological behavior by the bacterial interaction. This study showed that hydrogen peroxide generated by redox-active compound derived from a breakdown product of siderophore can significantly increase the number of hyphae tip extension in filamentous bacteria. Our result implies the existence of oxidative response system using a low amount of reactive oxygen species as an integrated signal to sense the plant-derived carbon source by the filamentous soil bacteria. As a result of sensing, filamentous soil bacteria may decide whether the hypha tip should be extended to further explore the area or increase the tips to densify filamentous network to monopolize the nutrients in the milieu.
    Cold Spring Harbor Laboratory, 2023年01月13日, bioRxiv

  • ミコール酸含有細菌の接触刺激による放線菌二次代謝応答機構の変異ゲノム解析を用いた解明
    浅水俊平
    2020年12月, Institute for Fermentation, Osaka. Research Communications, (34) (34)

  • 物理的接触が介在する放線菌二次代謝誘導
    尾仲宏康, 浅水俊平
    2020年06月15日, Journal of Environmental Biotechnology (Web), 20(1) (1)

  • 新規ラビオネン構造の形成に関与するランチペプチド合成酵素の解析—平成24年度寄付講座助成
    菅井 佳宣, 尾﨑 太郎, 浅水 俊平, 尾仲 宏康
    大阪 : 発酵研究所, 2018年12月20日, Research communications = 発酵研究所研究報告, (32) (32), 167 - 177, 日本語

  • ミコール酸含有細菌に対する放線菌二次代謝応答機構の網羅的転写解析—平成24年度寄付講座助成
    浅水 俊平, 尾﨑 太郎, 菅井 佳宣, 尾仲 宏康
    大阪 : 発酵研究所, 2018年12月20日, Research communications = 発酵研究所研究報告, (32) (32), 87 - 94, 日本語

  • 重イオンビーム変異導入法によるTsukamurella pulmonisへの赤色色素非生産応答性Streptomyces coelicolor突然変異株の解析—平成24年度寄付講座助成
    浅水 俊平, 菅井 佳宣, 尾﨑 太郎, 尾仲 宏康
    大阪 : 発酵研究所, 2018年12月20日, Research communications = 発酵研究所研究報告, (32) (32), 95 - 104, 日本語

  • ゴードスポリン作用機構の解析—平成24年度寄付講座助成
    尾﨑 太郎, 菅井 佳宜, 浅水 俊平, 尾仲 宏康
    大阪 : 発酵研究所, 2018年12月20日, Research communications = 発酵研究所研究報告, (32) (32), 129 - 139, 日本語

  • In vitro再構成系を用いたゴードスポリン生合成酵素の解析と合理的な類縁体設計—平成24年度寄付講座助成
    尾﨑 太郎, 菅井 佳宣, 浅水 俊平, 尾仲 宏康
    大阪 : 発酵研究所, 2018年12月20日, Research communications = 発酵研究所研究報告, (32) (32), 141 - 154, 日本語

  • ゲノム探索による32員環新規チオペプチドラクタゾールの発見—平成24年度寄付講座助成
    林 昌平, 尾﨑 太郎, 浅水 俊平, 尾仲 宏康
    大阪 : 発酵研究所, 2018年12月20日, Research communications = 発酵研究所研究報告, (32) (32), 155 - 166, 日本語

  • 放線菌複合培養液からの抗生物質-シデロフォア連結化合物の発見—平成24年度寄付講座助成
    浅水 俊平, 菅井 佳宣, 尾仲 宏康
    大阪 : 発酵研究所, 2018年12月20日, Research communications = 発酵研究所研究報告, (32) (32), 117 - 127, 日本語

  • ミコール酸含有細菌死菌体は放線菌の二次代謝を誘導しない—平成24年度寄付講座助成
    浅水 俊平, 尾﨑 太郎, 尾仲 宏康
    大阪 : 発酵研究所, 2018年12月20日, Research communications = 発酵研究所研究報告, (32) (32), 73 - 86, 日本語

  • 複合培養を用いた新規アルカロイドの同定と生合成に関する研究—平成24年度寄付講座助成
    尾﨑 太郎, 浅水 俊平, 尾仲 宏康
    大阪 : 発酵研究所, 2018年12月20日, Research communications = 発酵研究所研究報告, (32) (32), 105 - 115, 日本語

  • 異属間相互作用により誘導される放線菌の特殊代謝—特集 微生物の「声」が聴きたくて…単細胞生物のコミュニケーションスキル
    浅水 俊平, 尾仲 宏康
    コレクション : 国立国会図書館デジタルコレクション > 電子書籍・電子雑誌 > 学術機関 > 学協会
    吹田 : 日本生物工学会, 2018年, 生物工学会誌 / 日本生物工学会 編, 96(8) (8), 457 - 460, 日本語

  • Shumpei Asamizu
    Abstract Actinomycetes are a major source of bioactive natural products with important pharmaceutical properties. Understanding the natural enzymatic assembly of complex small molecules is important for rational metabolic pathway design to produce “artificial” natural products in bacterial cells. This review will highlight current research on the biosynthetic mechanisms of two classes of nitrogen-containing natural products, C7N aminocyclitols and bis-indoles. Validamycin A is a member of C7N aminocyclitol natural products from Streptomyces hygroscopicus. Here, two important biosynthetic steps, pseudoglycosyltranferase-catalyzed C–N bond formation, and C7-sugar phosphate cyclase-catalyzed divergent carbasugar formation, will be reviewed. In addition, the bis-indolic natural products indolocarbazole, staurosporine from Streptomyces sp. TP-A0274, and rearranged bis-indole violacein from Chromobacterium violaceum are reviewed including the oxidative course of the assembly pathway for the bis-indolic scaffold. The identified biosynthesis mechanisms will be useful to generating new biocatalytic tools and bioactive compounds.
    Informa UK Limited, 2017年05月04日, Bioscience, Biotechnology, and Biochemistry, 81(5) (5), 871 - 881
    [査読有り][招待有り]

  • Exploiting the potential of biosynthesis of natural products by actinomycetes: bacterial interaction-driven natural product discovery and biosynthetic machinery
    浅水 俊平
    2017年06月, Actinomycetologica, 31(1) (1), S30 - S40, 国際共著していない
    [招待有り]

  • もう無視できない天然物資源(バイオミディア)
    浅水,俊平
    日本生物工学会, 2015年08月25日, 生物工学会誌 : Seibutsu-kogaku Kaishi, 93(8) (8), 489, 日本語

■ 講演・口頭発表等
  • 複合培養における色素生産応答に関わるsco1842遺伝子の機能解析
    久保木綾梨, LEI Yukun, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2024年

  • De novo RiPPs創薬を志向した非天然チオペプチド発酵生産系の確立
    伊地知新太, 永井栄美子, 浅水俊平, VINOGRADOV Alexander A., 後藤佑樹, 菅裕明, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2024年

  • 細菌細胞内に局在するファージ尾部様粒子が有する推定エフェクタータンパク質の同定
    永久保利紀, 西山辰也, 浅水俊平, 尾仲宏康, 野村暢彦, 豊福雅典
    日本農芸化学会大会講演要旨集(Web), 2024年

  • 放線菌におけるsigRを介した酸化ストレス応答のピロガロール誘導型分岐形成への関与
    福原彩穂, 加藤愛美, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2024年

  • RiPP創薬を志向する非天然環状ペプチドの発酵生産
    伊地知新太, 永井栄美子, 星野翔太郎, 浅水俊平, VINOGRADOV Alexander A., 後藤佑樹, 後藤佑樹, 菅裕明, 尾仲宏康
    日本生物工学会大会講演要旨集, 2024年

  • 宿主細胞のストレス応答に関連づけられたファージ尾部様粒子の発見とその生態学的影響の示唆
    永久保利紀, 西山辰也, 浅水俊平, 山本達也, 加藤愛美, 野村暢彦, 豊福雅典, 尾仲宏康
    日本微生物生態学会大会(Web), 2023年11月

  • 自然環境からのシグナル:ピロガロールを介した放線菌の分枝形成と環境応答について
    加藤愛美, 浅水俊平, 尾仲宏康
    日本微生物生態学会大会(Web), 2023年11月

  • 微生物間化学コミュニケーションが促進する放線菌の分枝応答
    浅水俊平, 加藤愛美, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2023年09月

  • 放線菌に分布する有機ヒ素二次代謝経路の開拓
    星野翔太郎, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2023年09月

  • Labioninへの酸素添加は抗菌活性を向上させる
    伊地知新太, 星野翔太郎, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2023年09月

  • Streptomyces属の二次代謝に関与する属内高度保存遺伝子の解析
    LEI Yukun, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2023年03月

  • 新規solabionin骨格の形成に関わる酸化還元酵素の解析
    伊地知新太, 星野翔太郎, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2023年03月

  • 放線菌に保存されたファージ尾部様粒子の機能および局在解析
    永久保利紀, 浅水俊平, 山本達也, 加藤愛美, 豊福雅典, 野村暢彦, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2023年03月

  • 放線菌におけるヒ素二次代謝経路に関する研究
    星野翔太郎, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2023年03月

  • セカンドメッセンジャー・c-di-GMPを介した放線菌薬物排出機構の解析
    LEI Yukun, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2022年09月

  • 同位体取込みを指標としたメタボロミクスによる極性官能基含有脂肪酸含有天然物の発見
    浅水俊平, 星野翔太郎, 伊地知新太, JO Hanseam, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2022年09月

  • 放線菌が生産する有機ヒ素二次代謝産物に関する研究
    星野翔太郎, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2022年09月

  • 高度に保存されたウイルス関連遺伝子群に着目した放線菌-異種微生物間相互作用の解析
    永久保利紀, 山本達也, 浅水俊平, 豊福雅典, 野村暢彦, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2022年09月

  • RiPPs前駆体遺伝子の転写ターミネーターに注目したチオペプチド異種生産系の検討
    伊地知新太, 永井栄美子, 星野翔太郎, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2022年09月

  • 土壌微生物と共存する新規ウイルス様粒子の生物学的意義
    永久保利紀, 浅水俊平, 山本達也, 豊福雅典, 野村暢彦, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2022年03月

  • 放線菌に対して生育阻害や二次代謝・胞子形成誘導を起こすペプチド系抗生物質goadsporinの作用機構解析
    河野佐知子, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2022年03月

  • セカンドメッセンジャー・c-di-GMPを介した放線菌薬物排出機構の発見
    LEI Yukun, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2022年03月

  • 新規な放線菌分枝誘導物質の発見
    加藤愛美, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2022年03月

  • 新規リポランチンの単離同定
    JO Hansaem, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2022年03月

  • 放線菌の二次代謝を中心に見た土壌微生物の関わり合い
    浅水俊平
    日本農芸化学会中部支部例会講演要旨集(Web), 2021年11月

  • 複合培養におけるStreptomyces coelicolorのRED生産遅延に関与するTetR型転写制御因子の解析
    LEI Yukun, 浅水俊平, 石塚匠, 柳澤昌臣, 大野豊, 佐藤勝也, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2021年03月

  • ゴードスポリンがStreptomyces lividansの生産するタンパク質及び生育,二次代謝に与える影響
    河野佐知子, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2021年03月

  • 放線菌にコブ状形態を誘導するピロガロールの解析
    加藤愛美, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2021年03月

  • goadvioninにおける脂肪酸部位の生合成の解析
    小境陵介, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2021年03月

  • 薬剤排出様ABCトランスポーターの高発現による複合培養における二次代謝生産応答の遅延機構の解析
    LEI Yukun, 浅水俊平, 石塚匠, 柳澤昌臣, 大野豊, 佐藤勝也, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2021年09月

  • 放線菌に広く保存されたファージ尾部様粒子のイメージング解析
    永久保利紀, 浅水俊平, 豊福雅典, 野村暢彦, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2021年09月

  • 枯草菌とミコール酸含有細菌によって共生産されるフェノール性化合物が放線菌の分枝を促進する
    加藤愛美, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2021年09月

  • 放線菌が生産する新規リポランチン類の探索
    浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2021年09月

  • 新規リボソーム翻訳型ペプチド/ポリケタイドハイブリッド化合物・ゴードビオニンの発見とその生合成
    小野拓人, 星野翔太郎, 小境陵介, 高橋秀典, 寺本華奈江, 田中耕一, 阿部郁朗, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2021年09月

  • 放線菌が生産する擬似糖鎖の網羅的生合成解析
    角田毅, 伊藤卓也, 浅水俊平, SAMUEL Tanoeyadi, TAIFO Mahmud
    日本放線菌学会大会講演要旨集, 2021年09月

  • 放線菌に対して生育阻害や二次代謝・胞子形成誘導を起こすペプチド系抗生物質goadsporinの作用機構解析
    河野佐知子, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2021年09月

  • RNA-seq解析に基づいた複合培養における二次代謝活性化機構の解析
    木田優士, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2021年03月

  • Streptomyces sp.HEK616とTsukamurella pulmonisとの複合培養での二次代謝活性化機構の解析
    浅水俊平, 尾崎太郎, 西村慎一, 掛谷秀昭, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2019年09月

  • 自然共分離放線菌とミコール酸含有細菌の生態学的相互作用の解析
    加藤愛美, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2019年09月

  • GNAT superfamily,GdvGの広い基質認識を利用した新規リポペプチド創製
    小境陵介, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2019年09月

  • SRP(シグナル認識粒子)を分子標的とした新規抗生物質探索系の確立
    永井栄美子, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2019年09月

  • 放線菌の形態異常を引き起こす三種複合培養の解析
    加藤愛美, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2019年03月

  • 複合培養におけるStreptomyces sp.HEK616による5aTHQの生産機構
    浅水俊平, 尾崎太郎, 西村慎一, 掛谷秀昭, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2019年03月

  • 複合培養における赤色色素生産非応答性変異株の解析
    石塚匠, 浅水俊平, 柳澤昌臣, 佐藤勝也, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2019年03月

  • 複合培養により活性化される二次代謝遺伝子プロモーターの制御機構の解析
    桐原正隆, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2019年03月

  • N-アセチルトランスフェラーゼ様酵素がポリケチドとランチペプチドの新規結合反応を触媒する
    小境陵介, 菅井佳宣, 寺本和矢, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2019年03月

  • ミコール酸含有細菌に対する二次代謝非応答性放線菌変異株の変異点の同定
    浅水俊平, 石塚匠, 柳澤昌臣, 寺本和矢, 佐藤勝也, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2018年09月

  • godAプロモーターの複合培養に応答した転写活性化機構の解析
    桐原正隆, 浅水俊平, 寺本和矢, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2018年09月

  • 自然共分離株Streptomyces variegatusとMycobacterium septicumの接触相互作用の解析
    加藤愛美, 浅水俊平, 寺本和矢, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2018年09月

  • Streptomyces sp.HOK021とTsukamurella pulmonisの組合せ培養からの新しい抗生シデロホアハイブリッド化合物の発見
    PRAMANA Abrory Agus Cahya, KAWAI Sungjin, KATO Manami, ASAMIZU Shumpei, SUGAI Yoshinori, SUZUKI Hiroaki, ARAKAWA Yoshichika, ONAKA Hiroyasu
    日本農芸化学会大会講演要旨集(Web), 2018年03月

  • 異属細菌間の相互作用がもたらす放線菌特殊代謝
    浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2018年03月

  • 舳倉島自然土壌から共分離された放線菌とミコール酸含有細菌の相互作用
    加藤愛美, 浅水俊平, 菅井佳宣, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2018年03月

  • 二次代謝誘導ペプチドgoadsporinの作用機構
    金田彬, 尾崎太郎, 菅井佳宣, 浅水俊平, 千田俊哉, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2018年03月

  • 新規labionen構造の形成に関与するlanthipeptide合成酵素の解析
    菅井佳宣, 江畑一真, 浅水俊平, 後藤佑樹, 菅裕明, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2018年03月

  • 複合培養法を利用した希少放線菌からの生物活性物質探索
    星野翔太郎, 淡川孝義, 浅水俊平, 尾仲宏康, 阿部郁朗
    日本薬学会年会要旨集(CD-ROM), 2018年

  • 微生物の複合培養で得られる5aTHQの膜親和性と生物活性
    西村慎一, 西村慎一, 杉山龍介, 仲谷崇宏, 尾崎太郎, 浅水俊平, 尾仲宏康, 掛谷秀昭
    天然有機化合物討論会講演要旨集(Web), 2018年

  • 自然土壌から共分離された放線菌とミコール酸含有細菌の相互作用の解析
    加藤愛美, 浅水俊平, 菅井佳宣, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2017年09月

  • 複合培養によって生産されるテトラヒドロキノリン化合物の解析
    下村杜人, 尾崎太郎, 杉山龍介, 西村慎一, 浅水俊平, 掛谷秀昭, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2017年09月

  • Goadsporinとそのターゲットタンパク質Ffhの相互作用の解析
    金田彬, 菅井佳宣, 浅水俊平, 千田俊哉, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2017年09月

  • RiPP-polyketideハイブリッド化合物の生合成に関与するGNAT型アシル基転移酵素の解析
    菅井佳宣, 千田美紀, 小野拓人, 浅水俊平, 千田俊哉, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2017年09月

  • ミコール酸含有細菌に対する放線菌の網羅的転写応答解析
    浅水俊平, 菅井佳宣, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2017年03月

  • 複合培養における放線菌の二次代謝活性化に関与する遺伝子のイオンビームを用いた逆遺伝学的解析
    柳澤昌臣, 浅水俊平, 菅井佳宣, 佐藤勝也, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2017年03月

  • 新規RiPPs-ポリケタイドハイブリッド化合物の生合成経路解析
    菅井佳宣, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2017年03月

  • 薬剤ターゲットをFfhとするゴードスポリンの新規作用機構解析
    金田彬, 尾崎太郎, 菅井佳宣, 浅水俊平, 千田俊哉, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2017年03月

  • 試験管内完全合成系によるゴードスポリン生合成経路の解析とデザイナーアナログ創製
    尾仲宏康, 尾崎太郎, 山下湖奈, 後藤佑樹, 下村杜人, 林昌平, 浅水俊平, 菅井佳宣, 菅裕明
    日本農芸化学会大会講演要旨集(Web), 2017年03月

  • 放線菌の眠れる二次代謝を呼び覚ます!-異属細菌間での競合と協調-
    浅水俊平
    日本生物工学会大会講演要旨集, 2017年

  • 複合培養法を用いた希少放線菌からの医薬品資源探索
    星野翔太郎, 淡川孝義, 岡田正弘, 浅水俊平, 尾仲宏康, 阿部郁朗
    食品薬学シンポジウム講演要旨集, 2017年

  • 重イオンビーム遺伝子変異導入法を用いた複合培養による放線菌の二次代謝活性化に関与する遺伝子の探索
    柳澤昌臣, 浅水俊平, 菅井佳宣, 佐藤勝也, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2016年09月

  • 複合培養におけるStreptomyces coelicolor A3(2)の経時的トランスクリプトーム解析
    浅水俊平, 佐々木槇子, 尾崎太郎, 菅井佳宣, 宮本憲二, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2016年09月

  • 放線菌由来の窒素含有天然物生合成に関する研究
    浅水俊平
    日本放線菌学会大会講演要旨集, 2016年09月

  • 新規ポリケタイド-RiPPsハイブリッド化合物の生合成研究
    菅井佳宣, 小野拓人, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2016年09月

  • 無細胞翻訳後系を利用したゴードスポリン生合成経路の再構成と新奇アナログの創製
    尾崎太郎, 山下湖奈, 下村杜人, 後藤佑樹, 菅井佳宣, 浅水俊平, 菅裕明, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2016年09月

  • 無細胞翻訳系を用いたラクタゾールの試験管内再構成系の確立
    下村杜人, 尾崎太郎, 菅井佳宣, 浅水俊平, 山下湖奈, 後藤佑樹, 菅裕明, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2016年09月

  • 複合培養特異的に生産されるテトラヒドロキノリン生合成遺伝子の解析
    下村杜人, 尾崎太郎, 杉山龍介, 西村慎一, 浅水俊平, 掛谷秀昭, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2016年03月

  • RNA-seqによるミコール酸含有細菌に対するStreptomyces coelicolor A3(2)の応答解析
    佐々木槇子, 浅水俊平, 尾崎太郎, 宮本憲二, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2016年03月

  • 二次代謝誘導物質ゴードスポリン作用機構の解析
    尾崎太郎, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2016年03月

  • ゴードスポリンの酵素的全合成
    山下湖奈, 尾崎太郎, 浅水俊平, 後藤佑樹, 菅裕明, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2016年03月

  • 脂質結合性天然物の活用-放線菌複合培養液より見出した新規アルカロイド群に関する研究
    杉山龍介, 西村慎一, 尾崎太郎, 浅水俊平, 尾仲宏康, 掛谷秀昭
    日本薬学会年会要旨集(CD-ROM), 2016年

  • 微生物の複合培養で得られる5aTHQとstreptoaminalの構造と機能
    西村慎一, 杉山龍介, 仲谷崇宏, 尾崎太郎, 浅水俊平, 尾仲宏康, 掛谷秀昭
    天然薬物の開発と応用シンポジウム講演要旨集, 2016年

  • 放線菌由来窒素含有天然生物活性物質の生合成に関する研究
    浅水俊平
    日本農芸化学会関東支部講演要旨集, 2016年

  • 複合培養における放線菌とミコール酸含有菌の生死で異なる相互作用
    浅水俊平, 尾崎太郎, 寺本華奈江, 佐藤勝也, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2015年09月

  • 二次代謝誘導物質ゴードスポリン作用機構の解析
    尾崎太郎, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2015年09月

  • Streptomyces sp.TP-A0584由来ゴードスポリン生合成の試験管内再構成系の確立
    山下湖奈, 尾崎太郎, 浅水俊平, 後藤佑樹, 菅裕明, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2015年09月

  • 二次代謝誘導物質ゴードスポリンの作用機構の解析
    尾崎太郎, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2015年03月

  • goadsporin生合成機構を利用したgoadsporin倍加ペプチドの生産
    岡本悠志, 尾崎太郎, 浅水俊平, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2015年03月

  • 複合培養における放線菌二次代謝の誘導要因の探索
    浅水俊平, 尾崎太郎, 佐藤勝也, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2015年03月

  • Streptomyces sp.TP-A0584株由来ゴードスポリン生合成のin vitro再構成
    山下湖奈, 尾崎太郎, 浅水俊平, 後藤佑樹, 菅裕明, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2015年03月

  • Streptomyces sp.TP-A0584由来ランチペプチド系化合物のゲノムマイニングによる同定
    小野拓人, 江畑一真, 浅水俊平, 尾崎太郎, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2015年03月

  • 複合培養を利用した新規テトラヒドロキノリン類の単離とその生合成に関する研究
    尾崎太郎, 杉山龍介, 西村慎一, 浅水俊平, 掛谷秀昭, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2014年09月

  • 死細胞は放線菌の二次代謝を複合培養において誘導しない
    浅水俊平, 尾崎太郎, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2014年09月

  • 複合培養において死細胞は放線菌の二次代謝を誘導しない
    浅水俊平, 尾崎太郎, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2014年03月

  • 放線菌の複合培養法による新規テトラヒドロキノリン類の単離とその生合成
    尾崎太郎, 森夕希子, 杉山龍介, 西村慎一, 浅水俊平, 掛谷秀昭, 尾仲宏康
    日本農芸化学会大会講演要旨集(Web), 2014年03月

  • ゲノム探索が明らかにした新規32員環チオペプチド,ラクタゾールの構造と生合成
    林昌平, 尾崎太郎, 浅水俊平, 尾仲宏康
    天然有機化合物討論会講演要旨集(Web), 2014年

  • 異属細菌による放線菌二次代謝産物の誘導
    浅水俊平, 尾仲宏康
    日本農芸化学会関東支部講演要旨集, 2014年

  • 二次代謝誘導物質ゴードスポリン耐性化機構の解析
    尾崎太郎, 浅水俊平, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2013年

  • 複合培養に応答するgodAプロモーター領域の解析
    浅水俊平, 小野拓人, 林昌平, 尾崎太郎, 尾仲宏康
    日本放線菌学会大会講演要旨集, 2013年

■ 所属学協会
  • 日本放線菌学会
    2013年04月 - 現在

  • 日本農芸化学会
    2013年04月 - 現在

■ 共同研究・競争的資金等の研究課題
  • 放線菌の分枝発生メカニズムの解析と応用
    公益財団法人発酵研究所（IFO）, 2024年04月 - 2026年03月

  • 酵素プロミスキュイティと生合成工学的手法を用いた非天然 RiPP型リポペプチドの生合成
    2024年度 ノボザイムズ ジャパン研究ファンド, 2024年01月 - 2024年12月

  • 浅水 俊平
    日本学術振興会, 科学研究費助成事業 基盤研究(C), 基盤研究(C), 東京大学, 2018年04月01日 - 2021年03月31日
    シデロマイシンはシデロフォアと抗生物質が連結した化合物である。放線菌Streptomyces sp. HOK021はミコール酸含有細菌Tsukamurella pulmonis TP-B0596との複合培養特異的に、脂肪酸合成酵素阻害剤であるプラテンシマイシンとカテコール型シデロフォアであるエンテロバクチン様構造がチオエステル架橋によって連結したシデロマイシン様キメラ抗生物質を蓄積した。本研究では、生産菌のゲノム解析からHDMの生合成機構に関する知見、既存のシデロマイシン類の質量分析における物理化学特性に関する知見を得た。新規シデロマイシン系抗生物質の探索研究への応用が期待できる。

  • ミコール酸含有細菌の接触刺激による放線菌二次代謝応答機構の変異ゲノム解析を用いた解明
    公益財団法人発酵研究所（IFO）, 2018年04月 - 2020年03月, 研究代表者

  • 放線菌由来シデロマイシン系抗生物質の生合成及び作用機作解析
    公益財団法人野田産業科学研究所, 2018年04月 - 2019年03月, 研究代表者

  • 浅水 俊平
    日本学術振興会, 科学研究費助成事業 若手研究(B), 若手研究(B), 東京大学, 2016年04月01日 - 2018年03月31日
    放線菌は医農薬として利用される有用天然物の生産者であり、近年のゲノム解析から一菌株当り20以上の有用天然物を生産する能力があることが示唆されている。本課題ではミコール酸含有細菌による放線菌休眠二次代謝遺伝子の活性化機構に関わる遺伝子の同定を目的に研究を行った。共培養時の網羅的転写再解析の結果、放線菌転写変動遺伝子の数は8時間で全遺伝子の約2割にも及んだ。また、非応答性の放線菌変異株スクリーニングを行い、現在16株のゲノムリシーケンスからいくつかの二次代謝産物生産に関わる変異遺伝子の同定に至った。今後の更なる解析により活性化の分子機構を明らかにすることを現在目指している。

  • 浅水 俊平
    日本学術振興会, 科学研究費助成事業 若手研究(B), 若手研究(B), 東京大学, 2014年04月 - 2016年03月
    放線菌の二次代謝産物は医薬・農薬として利用されている。放線菌ゲノム解析から潜在的二次代謝遺伝子群が一菌株に20～40個存在する明らかになっている。本研究では放線菌に眠っている二次代謝遺伝子を覚醒させる手法として、放線菌とミコール酸含有細菌との共培養（複合培養）法に注目した。本課題では二次代謝遺伝子の活性化機構を明らかにするために、生産量が増大することが明らかになったゴードスポリンの生合成遺伝子に注目し、前駆体ペプチド遺伝子の発現プロモーターを明らかにした。この結果と網羅的転写解析の結果を考え合わせ、二次代謝産物の生産量の増加には細胞全体の一次代謝変化が一因となっていると考えられた。

  • ミコール酸含有細菌による放線菌二次代謝活性化機構に関する研究
    公益信託荒木記念医学・生化学研究振興基金, 2014年04月 - 2015年03月, 研究代表者

  • インドロカルバゾール生合成機構の解明及び新規類縁体のコンビナトリアル生合成
    浅水 俊平
    日本学術振興会, 科学研究費助成事業 特別研究員奨励費, 特別研究員奨励費, 富山県立大学, 2008年 - 2009年
    本研究は、微生物が生産する抗癌活性を有するビスインドール化合物に注目し、一つ目に、ビスインドール化合物生合成酵素のX線結晶構造に基づいたアミノ酸置換による新たな酵素機能の開発を目的とし、二つ目に、改変酵素を用いたビスインドール新規類縁体のコンビナトリアル生合成による環境負荷に配慮した生産法の確立を目的としている。これまでに、ビスインドール化合物の詳細な生合成機構を解明するために、スタウロスポリン生合成酵素StaPとビオラセイン生合成酵素VioEのX線結晶構造を明らかにし、StaPとVioEが担う詳細な反応機構を明らかにしている。一方で、明らかになったStaPとVioE反応機構から、アミノ酸置換により、それぞれの酵素反応を改変することは困難であることも示唆された。そこで本年度は、RebCのX線結晶構造に基づき、新たにスタウロスポリン生合成酵素StaCのアミノ酸置換を行い、in vitroでStaC酵素反応を改変することに成功した。StaCはStaPと協調してクロモピロリン酸からK252cを合成するモノオキシゲナーゼホモログである。本酵素の反応機構は未解明な部分が多いが、RebCのX線結晶構造中で基質ポケットを構成する二か所のアミノ酸残基をStaCに導入したところ、RebC型の活性と共に、抗癌剤として臨床開発試験中であるUCN-01の基本骨格である7-hydroxy-K252cを酵素的に生成することを見出した。UCN-01の有効な培養生産法はこれまでになかったことから、本改変StaC酵素を用いることで培養生産が改善出来る可能性が見出された。また、更にインドロカルバゾール化合物の骨格構造の多様性に貢献できると考えられる。
    競争的資金

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