KURATA Atsushi
Department of Applied Biological Chemistry | Professor |
Last Updated :2024/09/14
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Research Keyword
- 乳酸菌 ビフィズス菌 酢酸菌 納豆菌 細胞外膜小胞 アポトーシス 免疫賦活作用
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Award
- 2024/03 Japan Society for Bioscience, Biotechnology, and Agrochemistry Award for Excellence to Authors Publishing in Bioscience, Biotechnology, and Biochemistry in 2023
Enhancement of IgA production by membrane vesicles derived from Bifidobacterium longum subsp. infantis受賞者: Atsushi Kurata;Shino Yamasaki-Yashiki;Tomoya Imai;Ayano Miyazaki;Keito Watanabe;Koichi Uegaki
Paper
- Atsushi Kurata; Kota Aimatsu; Yuki Kimura; Hinako Hashiguchi; Asami Maeda; Tomoya Imai; Shino Yamasaki-Yashiki; Kensaku Hamada; Yuki Fujimoto; Akira Fujii; Koichi UegakiJournal of Bioscience and Bioengineering Elsevier BV 1389-1723 2024/09
- Atsushi Kurata; Shimpei Takeuchi; Ryo Fujiwara; Kento Tamura; Tomoya Imai; Shino Yamasaki-Yashiki; Hiroki Onuma; Yasuhisa Fukuta; Norifumi Shirasaka; Koichi UegakiBioscience, Biotechnology, and Biochemistry Oxford University Press (OUP) 87 (8) 907 - 915 1343-327X 2023/05 [Refereed]
ABSTRACT We characterized the membrane vesicle fraction (RD-MV fraction) from bacterial strain RD055328, which is related to members of the genus Companilactobacillus and Lactiplantibacillus plantarum. RD-MVs and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were detected in the RD-MV fraction. Immunoglobulin A (IgA) was produced by Peyer's patch cells following the addition of the RD-MV fraction. In the presence of the RD-MV fraction, RAW264 cells produced the pro-inflammatory cytokine IL-6. Recombinant GAPDH probably induced the production of IL-6 by RAW264 cells via superficial toll-like receptor 2 (TLR2) recognition. A confocal laser scanning microscopy image analysis indicated that RD-MVs and GAPDH were taken up by RAW264 cells. GAPDH wrapped around RAW264 cells. We suggest that GAPDH from strain RD055328 enhanced the production of IgA by acquired immune cells via the production of IL-6 by innate immune cells through TLR2 signal transduction. - 志岐 幸祐; 立石 里佳; 藤崎 明日香; 倉田 淳志; 山田 秀和; 水口 信行; 伊藤 龍生日本臨床腸内微生物学会誌 25 (1) 38 - 45 2023/03 [Refereed]
- Atsushi Kurata; Shino Yamasaki-Yashiki; Tomoya Imai; Ayano Miyazaki; Keito Watanabe; Koichi UegakiBioscience, biotechnology, and biochemistry 87 (1) 119 - 128 2022/12 [Refereed]
Immunoglobulin A (IgA) is involved in the maintenance of gut homeostasis. Although the oral administration of bifidobacteria increases the amount of fecal IgA, the effects of bifidobacteria on intestinal immunity remain unclear. We found and characterized membrane vesicles (MVs) derived from Bifidobacterium longum subsp. infantis toward host immune cells. Bifidobacterium infantis MVs consisted of a cytoplasmic membrane, and extracellular solute-binding protein (ESBP) was specifically detected. In the presence of B. infantis MVs or recombinant ESBP, RAW264 cells produced the pro-inflammatory cytokine IL-6. IgA was produced by Peyer's patches cells following the addition of B. infantis MVs. Therefore, ESBP of B. infantis MVs is involved in the production of IgA by acquired immune cells via the production of IL-6 by innate immune cells. - 倉田淳志日本乳酸菌学会誌 33 (3) 179 - 185 1343-327X 2022/11 [Invited]
- Atsushi Kurata; Shogo Kiyohara; Tomoya Imai; Shino Yamasaki-Yashiki; Nobuhiro Zaima; Tatsuya Moriyama; Noriaki Kishimoto; Koichi UegakiScientific reports 12 (1) 13330 - 13330 2022/08 [Refereed]
We investigated the characteristics and functionalities of extracellular vesicles (EVs) from Lactiplantibacillus plantarum (previously Lactobacillus plantarum) towards host immune cells. L. plantarum produces EVs that have a cytoplasmic membrane and contain cytoplasmic metabolites, membrane and cytoplasmic proteins, and small RNAs, but not bacterial cell wall components, namely, lipoteichoic acid and peptidoglycan. In the presence of L. plantarum EVs, Raw264 cells inducibly produced the pro-inflammatory cytokines IL-1β and IL-6, the anti-inflammatory cytokine IL-10, and IF-γ and IL-12, which are involved in the differentiation of naive T-helper cells into T-helper type 1 cells. IgA was produced by PP cells following the addition of EVs. Therefore, L. plantarum EVs activated innate and acquired immune responses. L. plantarum EVs are recognized by Toll-like receptor 2 (TLR2), which activates NF-κB, but not by other TLRs or NOD-like receptors. N-acylated peptides from lipoprotein19180 (Lp19180) in L. plantarum EVs were identified as novel TLR2 ligands. Therefore, L. plantarum induces an immunostimulation though the TLR2 recognition of the N-acylated amino acid moiety of Lp19180 in EVs. Additionally, we detected a large amount of EVs in the rat gastrointestinal tract for the first time, suggesting that EVs released by probiotics function as a modulator of intestinal immunity. - Hironobu Takagi; Kazuki Yamamoto; Yoshifumi Matsuo; Miki Furuie; Yasuha Kasayuki; Rina Ohtani; Mizuki Shiotani; Tetsuya Hasegawa; Toru Ohnishi; Masataka Ohashi; Katsuki Johzuka; Atsushi Kurata; Koichi UegakiBioscience, biotechnology, and biochemistry 86 (6) 755 - 762 2022/05Isoamyl alcohol (i-AmOH) is produced from α-ketoisocaproate in the l-leucine biosynthetic pathway in yeast and controlled by the negative feedback regulation of α-isopropylmalate synthase (IPMS), which senses the accumulation of l-leucine. It is known that i-AmOH production increases when mutations in the regulatory domain reduce the susceptibility to feedback inhibition. However, the impact of mutations in this domain on the IPMS activity has not been examined. In this study, we obtained 5 IPMS mutants, encoding the LEU4 gene, N515D/S520P/S542F/A551D/A551V, that are tolerant to 5,5,5-trifluoro-dl-leucine. All mutant proteins were purified and examined for both IPMS activity and negative feedback activity by in vitro experiments. The results showed that not only the negative-feedback regulation by l-leucine was almost lost in all mutants, but also the IPMS activity was greatly decreased and the difference in IPMS activity among Leu4 mutants in the presence of l-leucine was significantly correlated with i-AmOH production.
- 深海底泥からの有用細菌の探索、分類、生産物の特性解明倉田淳志2021/12 [Invited]
- Atsushi Kurata; Daichi Aoki; Yoshihumi Fukuta; Taichi Kamimura; Taiki Onishi; Noriaki Kishimoto; Koichi UegakiBiotechnology & Biotechnological Equipment 35 (1) 445 - 452 2021/02 [Refereed]
- Atsushi Kurata; Taishi Yamaguchi; Masaaki Kira; Noriaki KishimotoBiotechnology & Biotechnological Equipment Informa UK Limited 33 (1) 886 - 893 1310-2818 2019/01 [Refereed]
- Atsushi Kurata; Yuto Yamaura; Takumi Tanaka; Chiaki Kato; Kaoru Nakasone; Noriaki KishimotoWorld Journal of Microbiology and Biotechnology 33 (73) 1 - 8 2017/03 [Refereed]
- Atsushi Kurata; Miwa Sugiura; Kento Kokoda; Hiroaki Tsujimoto; Tetsuya Numata; Chiaki Kato; Kaoru Nakasone; Noriaki KishimotoBIOTECHNOLOGY & BIOTECHNOLOGICAL EQUIPMENT TAYLOR & FRANCIS LTD 31 (5) 1000 - 1006 1310-2818 2017 [Refereed]
Fifteen actinomycetes were isolated from deep-sea sediments in Calyptogena communities in the Japan Trench (depth: 5352m) and Sagami Bay (depth: 1171m) under incubation at 4-37 degrees C. A relatively large number of colonies were obtained with incubation at 30 degrees C from the deep-sea sediment of Sagami Bay. We found Rhodococcus, Nocardioides and Micromonospora actinomycetes in both sediments, and additionally isolated an Aeromicrobium actinomycete from the Japan Trench and Streptomyces and Actinomadura actinomycetes from Sagami Bay. Actinomadura sp. DS-MS-114 isolated from the deep-sea sediment in Sagami Bay produced an antibacterial compound against Staphylococcus aureus NBRC12732(T). Structural analysis by liquid chromatography-high-resolution electrospray ionization mass spectrometry (LC-HRESIMS) and proton nuclear magnetic resonance (H-1-NMR) indicated that the antibacterial compound was 5,6-dihydro-1,8-dihydroxy-3-methylbenz[a]anthracene-7,12-quinone, which is known as an immunosuppressant. The deep-sediment in the Calyptogena community may be useful as a source of various actinomycetes producing physiologically active substances. - Atsushi Kurata; Shota Shimizu; Yutaro Shiraishi; Mihoko Abe; Nobutoshi Naito; Mihoko Shimada; Noriaki KishimotoBIOTECHNOLOGY & BIOTECHNOLOGICAL EQUIPMENT TAYLOR & FRANCIS LTD 31 (4) 749 - 755 1310-2818 2017 [Refereed]
We demonstrated the degradation of two ionic liquids (1-butyl-3-methylimidazolium chloride, [BMIM]Cl, and 1-ethylpyridinium bromide, [EtPy]Br) that are useful for the solubilization of wood components. [BMIM](+) and [EtPy](+) were detected by thin-layer chromatography (TLC) and electrospray ionization-mass spectrometry (ESI-MS). [BMIM](+) was harder to degrade than [EtPy](+). Ultraviolet (UV) irradiation with 0.2% (v/v) H2O2 for 16 h degraded 1mmol/L [BMIM](+), whereas UV irradiation alone degraded 1mmol/L [EtPy](+). Additionally, we isolated an ionic liquid-tolerant alkaliphilic actinomycete, Nocardiopsis sp. SSC4. Strain SSC4 produced carboxymethylcellulase (CMCase) in the presence of 1.0% (v/v, 48.1mmol/L) 1-ethyl-3-methylimidazolium trifluoromethanesulphonate ([EMIM]CF3SO3), which is useful for the extraction of cellulose-rich materials from wood. In the case of strain SSC4, CMCase was inducibly synthesized by more than 0.5% CMC. The addition of 0%-1.0% tryptone or 0%-2.0% yeast extract decreased the CMCase activity in a concentration-dependent manner. After cultivation of strain SSC4 with 1.0% (w/v) CMC medium (pH 9.0) for 48h at 37 degrees C, the culture supernatant exhibited CMCase activity at 0.03U/mg. The optimum reaction temperature of CMCase was 45 degrees C. CMCase was stable up to 37 degrees C for 20h incubation. The degradation characteristics of [BMIM](+) and [EtPy](+) and the activity of CMCase in the presence of [EMIM]CF3SO3 may be useful for the development of a bioconversion system for biomass resources. - Atsushi Kurata; Humiya Senoo; Yasuyuki Ikeda; Hideaki Kaida; Chiaki Matsuhara; Noriaki KishimotoEXTREMOPHILES SPRINGER JAPAN KK 20 (4) 415 - 424 1431-0651 2016/07 [Refereed]
An ionic liquid-tolerant bacterium, Bacillus amyloliquefaciens CMW1, was isolated from a Japanese fermented soybean paste. Strain CMW1 grew in the presence of 10 % (v/v) 1-butyl-3-methylimidazolium chloride ([BMIM]Cl), a commonly used ionic liquid. Additionally, strain CMW1 grew adequately in the presence of the hydrophilic ionic liquids 10 % (v/v) 1-ethyl-3-methylimidazolium trifluoromethanesulfonate ([EMIM]CF3SO3) or 2.5 % (v/v) 1-butyl-3-methylimidazolium trifluoromethanesulfonate ([BMIM]CF3SO3). Strain CMW1 produced an extracellular protease (BapIL) in the culture medium. BapIL was stable in the presence of 80 % (v/v) ionic liquids, [EMIM]CF3SO3, [BMIM]Cl, [BMIM]CF3SO3, 1-butyl-3-methylimidazolium tetrafluoroborate, 1-butyl-3-methylimidazolium hexafluorophosphate, and 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide, and functioned in 10 % (v/v) these ionic liquids. BapIL was stable at pH 4.0-12.6 or in 4004 mM NaCl solution, and exhibited activity in the presence of 50 % (v/v) hydrophilic or hydrophobic organic solvents. BapIL was completely inhibited by 1 mM PMSF and partially by 5 mM EDTA. BapIL belongs to the true subtilisins according to analysis of the deduced amino acid sequence. We showed that BapIL from the ionic liquid-tolerant B. amyloliquefaciens CMW1 exhibited tolerance to ionic liquid and halo, alkaline, and organic solvents. - A high-molecular-weight, alkaline, and thermostable β-1,4-xylanase of a subseafloor Microcella alkaliphilaKoki Kuramochi; Kohsuke Uchimura; Atsushi Kurata; Tohru Kobayashi; Yuu Hirose; Takeshi Miura; Noriaki Kishimoto; Ron Usami; Koki HorikoshiExtremophiles 20 471 - 478 2016/03 [Refereed]
- Atsushi Kurata; Yuu Hirose; Naomi Misawa; Sachiko Wakazuki; Noriaki Kishimoto; Tohru KobayashiJOURNAL OF BIOTECHNOLOGY ELSEVIER SCIENCE BV 221 32 - 33 0168-1656 2016/03 [Refereed]
Here we report the complete genome sequence of Microcella alkaliphila JAM-AC0309, which was newly isolated from the deep subseafloor core sediment from offshore of the Shimokita Peninsula of Japan. An array of genes related to utilization of xylan in this bacterium was identified by whole genome analysis. (C) 2016 Elsevier B.V. All rights reserved. - Atsushi Kurata; Yasuhisa Fukuta; Miho Mori; Noriaki Kishimoto; Norifumi ShirasakaGenome Announcements American Society for Microbiology 4 (3) e00505-16 2169-8287 2016 [Refereed]
Here, we report the draft genome sequence of Flammulina velutipes TR19, which was newly isolated from commercial strains in Japan. The genes related to fruiting body formation in the basidiomycete were identified by whole-genome analysis. - Atsushi Kurata; Mioko Matsumoto; Tohru Kobayashi; Shigeru Deguchi; Noriaki KishimotoMARINE BIOTECHNOLOGY SPRINGER 17 (3) 277 - 284 1436-2228 2015/06 [Refereed]
A hyaluronate lyase (BniHL) was purified to homogeneity from a culture of a deep-sea Bacillus niacin strain JAM F8. The molecular mass of purified BniHL was approximately 120 kDa. The purified enzyme degraded hyaluronan as well as chondroitin sulfates A and C by a beta-elimination mechanism. The optimal pH and temperature were around pH 6 and 45 A degrees C for hyaluronan degradation. The enzyme required optimally 2, 50, and 100 mM calcium ions for degradation of hyaluronan, chondroitin sulfate C, and chondroitin sulfate A, respectively. Calcium ions slightly increased the thermal stability of the enzyme. In a genome analysis of strain JAM F8, a BniHL coding gene was identified on the bases of the molecular mass and N-terminal and internal amino acid sequences. The gene consisted of 3411 nucleotides and coded 1136 amino acids. The deduced amino acid sequence showed the highest similarity to the hyaluronate lyase of a Bacillus sp. A50 with 89 % identity. - H. Ando; A. Kurata; N. KishimotoJOURNAL OF APPLIED MICROBIOLOGY WILEY-BLACKWELL 118 (4) 873 - 880 1364-5072 2015/04 [Refereed]
AimsTo evaluate the antimicrobial properties of the main Ginjo-flavour components of sake, volatile isoamyl acetate and isoamyl alcohol. Methods and ResultsVolatile isoamyl acetate and isoamyl alcohol both inhibited growth of the five yeast and 10 bacterial test strains. The minimum inhibitory dose and minimum bactericidal (fungicidal) dose of isoamyl acetate were higher than those of isoamyl alcohol. Escherichia coli and Acetobacter aceti were markedly sensitive to isoamyl acetate and isoamyl alcohol. In E.coli exposed to isoamyl acetate for 5h, changes in expression were noted in proteins involved in sugar metabolism (MalE, MglB, TalB and PtsI), tricarboxylic acid cycle (AceA, Pfl and AcnB) and protein synthesis (EF-Tu, EF-G, and GlyS). Expression of acid and alcohol stress-response proteins was altered in E.coli exposed to isoamyl acetate. Esterase activity was detected in E.coli, suggesting that isoamyl acetate was hydrolyzed to acetic acid and isoamyl alcohol. Acetic acid and isoamyl alcohol damaged E.coli cell membranes and inactivated membrane proteins, impairing respiration. ConclusionsVolatile isoamyl acetate and isoamyl alcohol were effective in inactivating various micro-organisms, and antimicrobial mechanism of volatile isoamyl acetate against E.coli was clarified based on proteome analysis. Significance and Impact of the StudyTo the best of our knowledge, this is the first report to examine the antimicrobial mechanism of volatile organic compound using proteome analysis combining two-dimensional difference gel electrophoresis with peptide mass fingerprinting. - Hiroki Umemoto; Keigo Sawada; Atsushi Kurata; Shohei Hamaguchi; Satoshi Tsukahara; Takashi Ishiguro; Noriaki KishimotoJOURNAL OF OLEO SCIENCE JAPAN OIL CHEMISTS SOC 63 (7) 671 - 679 1345-8957 2014/07 [Refereed]
A high-nervonic acid (cis-15-tetracosenoic acid, C-24:1, n-9)-producing filamentous fungus of the Mortierella species was discovered among soil filamentous fungi. The filamentous fungal strain -RD000969- was isolated from soil collected in Kanagawa Prefecture (Japan) and was found to accumulate nervonic acid at a rate of 6.94% of the total cellular fatty acids. The base sequences of 28S rDNA D1/D2 and ITS 5.8S rDNA showed 100% homology with Mortierella capitata CBS 293.96. In addition to nervonic acid, strain RD000969 produced a large amount of long-chain monounsaturated fatty acids (C-20:1, 12.22%; C-22:1, 4.07%; C-26:1, 5-91%) and a small amount of ultra-long-chain fatty acids (C-28:1, 0.44%; C-30:1, 0.06%; C-32:1, trace). In the fungal cells, 98.87% of nervonic acid was localized at the sn-1,3 position of triacylglycerol. Nervonic acid production was maximum (186.3 mg center dot L-1) when the fungus was cultured in potato dextrose (PD) medium containing yeast extract, CaCl2, and MgSO4 center dot 7H(2)O. - Atsushi Kurata; Yuu Hirose; Naomi Misawa; Kohei Hurunaka; Noriaki KishimotoGenome Announcements American Society for Microbiology 2 (5) e01051-14 2169-8287 2014 [Refereed]
Here, we report the draft genome sequence of an ionic liquid-tolerant bacterium, Bacillus amyloliquefaciens CMW1, which is newly isolated from a Japanese fermented soybean paste. The genome sequence will allow for a characterization of the molecular mechanism of its ionic liquid tolerance. - Atsushi Kurata; Midori Nishimura; Noriaki Kishimoto; Tohru KobayashiGenome Announcements American Society for Microbiology 2 (5) e00983-14 2169-8287 2014 [Refereed]
Here, we report the draft genome sequence of Bacillus niacini JAM F8, which was newly isolated from deep-sea sediment at a depth of 2,759mfrom the Izu-Ogasawara Trench. An array of genes related to degradation of glycosaminoglycans in this bacterium was identified by whole-genome analysis. - Hitoshi Ando; Koji Hatanaka; Ikumi Ohata; Yoshie Yamashita-Kitaguchi; Atsushi Kurata; Noriaki KishimotoFOOD CONTROL ELSEVIER SCI LTD 26 (2) 472 - 478 0956-7135 2012/08 [Refereed]
A yeast isolate, Candida maltosa NP9, was obtained from a fermented food, Iranian commercial cheese. The strain grown on YPD agar inhibited spore germination of Aspergillus brasiliensis by vapor-agar contact method. Seven volatile compounds generated from the strain were detected by SPME-GC/MS analysis, and three of them were identified as isoamyl alcohol, isoamyl acetate, and phenethyl alcohol by GC/MS analysis. Although phenethyl alcohol did not inhibit the germination at 160 mu l/dish by 48 h exposure, isoamyl alcohol fungicidally inhibited at 80 mu l/dish and isoamyl acetate fungiostatically inhibited at 160 mu l/dish. In antifungal spectra with 15 kinds of filamentous fungi, isoamyl alcohol inhibited the germination of all strains at 20 mu l/dish, while isoamyl acetate did not inhibit the germination of three strains even at 160 mu l/dish. We demonstrated that C. maltosa NP9 from the fermented food generated volatile isoamyl acetate and isoamyl alcohol to inhibit the germination of those fungi. (C) 2012 Elsevier Ltd. All rights reserved. - Anoop Vasudevan; Michiyo Fujita; Atsushi Kurata; Jun Kawamoto; Nobuyoshi Esaki; Tatsuo KuriharaTrace Nutrients Research 日本微量栄養素学会事務局 28 58 - 64 1346-2334 2011/10 [Refereed]
- Atsushi Kurata; Shintaro Takemoto; Tokio Fujita; Kazuya Iwai; Mina Furusawa; Noriaki KishimotoJOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC ELSEVIER SCIENCE BV 69 (3-4) 161 - 167 1381-1177 2011/05 [Refereed]
We developed a convenient one-pot procedure for conversion of 5-caffeoylquinic acid to 3-cyclohexylpropyl caffeate, which exhibits an antiproliferative effect toward various human tumor cells. The procedure was comprised of two consecutive reactions by chlorogenate hydrolase (EC 3.1.1.42) from Aspergillus japonicus and Candida antarctica lipase B, and was performed using an ionic liquid, 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide, as the reaction solvent. When various caffeoylquinic acids from coffee beans, namely, 3-caffeoylquinic acid, 4-caffeoylquinic acid, 5-caffeoylquinic acid,3,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid were used, the first alcoholysis reaction with methanol using chlorogenate hydrolase produced methyl caffeate with conversion yields of 60.0%, 61.3%, 86.0%, 92.7%, and 114.0%, respectively, to each individual substrate. Two caffeoyl groups of dicaffeoylquinic acids would be used for the synthesis of methyl caffeate. In the subsequent transesterification reaction by C. antarctica lipase B with 3-cyclohexyl-1-propanol, the methyl caffeate produced was converted to 3-cyclohexylpropyl caffeate under reduced pressure to remove the by-product methanol. In the one-pot synthesis, the methyl caffeate was transesterified efficiently to 3-cyclohexylpropyl caffeate by C. antarctica lipase B with deactivation of chlorogenate hydrolase by taking advantage of the difference between the optimum temperatures for the two enzymes. This system provided 12.8 mM 3-cyclohexylpropyl caffeate from 15 mM 5-caffeoylquinic acid with conversion yield of 85.3%. (C) 2011 Elsevier B.V. All rights reserved. - Amr M. Mowafy; Tatsuo Kurihara; Atsushi Kurata; Tadashi Uemura; Nobuyoshi EsakiAPPLIED AND ENVIRONMENTAL MICROBIOLOGY AMER SOC MICROBIOLOGY 76 (18) 6032 - 6037 0099-2240 2010/09 [Refereed]
Enzymes catalyzing the conversion of organohalogen compounds are useful in the chemical industry and environmental technology. Here we report the occurrence of a new reduced flavin adenine dinucleotide (FAD) (FADH(2))-dependent enzyme that catalyzes the removal of a halogen atom from an unsaturated aliphatic organohalogen compound by the addition of a water molecule to the substrate. A soil bacterium, Pseudomonas sp. strain YL, inducibly produced a protein named Caa67(YL) when the cells were grown on 2-chloroacrylate (2-CAA). The caa67(YL) gene encoded a protein of 547 amino acid residues (M(r) of 59,301), which shared weak but significant sequence similarity with various flavoenzymes and contained a nucleotide-binding motif. We found that 2-CAA is converted into pyruvate when the reaction was carried out with purified Caa67(YL) in the presence of FAD and a reducing agent [NAD(P) H or sodium dithionite] under anaerobic conditions. The reducing agent was not stoichiometrically consumed during this reaction, suggesting that FADH(2) is conserved by regeneration in the catalytic cycle. When the reaction was carried out in the presence of H(2) (18)O, [ (18)O] pyruvate was produced. These results indicate that Caa67(YL) catalyzes the hydration of 2-CAA to form 2-chloro-2-hydroxypropionate, which is chemically unstable and probably spontaneously dechlorinated to form pyruvate. 2-Bromoacrylate, but not other 2-CAA analogs such as acrylate and methacrylate, served as the substrate of Caa67(YL). Thus, we named this new enzyme 2-haloacrylate hydratase. The enzyme is very unusual in that it requires the reduced form of FAD for hydration, which involves no net change in the redox state of the coenzyme or substrate. - Atsushi Kurata; Yuki Kitamura; Shiori Irie; Shintaro Takemoto; Yoshiaki Akai; Yoshitaka Hirota; Tokio Fujita; Kazuya Iwai; Mina Furusawa; Noriaki KishimotoJOURNAL OF BIOTECHNOLOGY ELSEVIER SCIENCE BV 148 (2-3) 133 - 138 0168-1656 2010/07An efficient procedure for transesterification of methyl caffeate was developed to produce caffeic acid phenethyl ester analogues with Candida antarctica lipase B using an ionic liquid, 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide, as a solvent. The system provided 48.8 mM 2-cyclohexylethyl caffeate and 46.9 mM 3-cyclohexylpropyl caffeate with conversion yields of 97.6% and 93.8%, respectively. Reusability of the system was investigated, and the yield of 4-phenylbutyl caffeate was increased from 30.4 to 45.7 mM when the transesterification was carried out under reduced pressure to remove a by-product, methanol. Additionally, we showed that both 2-cyclohexylethyl caffeate and 3-cyclohexylpropyl caffeate exhibit strong antiproliferative activities, which are comparable to that of 5-fluorouracil by MTT assay. (C) 2010 Elsevier B.V. All rights reserved.
- Atsushi Kurata; Kohsuke Uchimura; Tohru Kobayashi; Koki HorikoshiAPPLIED MICROBIOLOGY AND BIOTECHNOLOGY SPRINGER 86 (2) 589 - 598 0175-7598 2010/03 [Refereed]
A new alkaline protease (AcpII) was purified from a culture of the deep-sea bacterium Alkalimonas collagenimarina AC40(T). AcpII degraded collagen three times faster than it degraded casein. The optimal pH was 8.5-9, and the optimal temperature was 45A degrees C for the degradation of collagen. AcpII was completely inhibited by phenylmethylsulfonyl fluoride and partially by EDTA. Cloning and sequencing the gene for AcpII revealed a 2,283-bp open reading frame encoding a protein of 760 amino acids. AcpII comprises a prepropeptide, a catalytic domain that includes a protease-associated domain (PA domain), and tandem repeat prepeptidase C-terminal domains. To elucidate the role of the PA domain of AcpII, we constructed genes for two enzyme derivatives that possessed the catalytic domains with or without the PA domain and expressed them in Escherichia coli. The derivative without the PA domain showed increased specific activities toward all proteinaceous substrates tested, including gelatin, casein, and collagen, compared with those of the derivative with the PA domain. - Keiji Jitsumori; Rie Omi; Tatsuo Kurihara; Atsushi Kurata; Hisaaki Mihara; Ikuko Miyahara; Ken Hirotsu; Nobuyoshi EsakiJOURNAL OF BACTERIOLOGY AMER SOC MICROBIOLOGY 191 (8) 2630 - 2637 0021-9193 2009/04 [Refereed]
Fluoroacetate dehalogenase catalyzes the hydrolytic defluorination of fluoroacetate to produce glycolate. The enzyme is unique in that it catalyzes the cleavage of a carbon-fluorine bond of an aliphatic compound: the bond energy of the carbon-fluorine bond is among the highest found in natural products. The enzyme also acts on chloroacetate, although much less efficiently. We here determined the X-ray crystal structure of the enzyme from Burkholderia sp. strain FA1 as the first experimentally determined three-dimensional structure of fluoroacetate dehalogenase. The enzyme belongs to the alpha/beta hydrolase superfamily and exists as a homodimer. Each subunit consists of core and cap domains. The catalytic triad, Asp104-His271-Asp128, of which Asp104 serves as the catalytic nucleophile, was found in the core domain at the domain interface. The active site was composed of Phe34, Asp104, Arg105, Arg108, Asp128, His271, and Phe272 of the core domain and Tyr147, His149, Trp150, and Tyr212 of the cap domain. An electron density peak corresponding to a chloride ion was found in the vicinity of the N-epsilon 1 atom of Trp150 and the N-epsilon 2 atom of His149, suggesting that these are the halide ion acceptors. Site-directed replacement of each of the active-site residues, except for Trp150, by Ala caused the total loss of the activity toward fluoroacetate and chloroacetate, whereas the replacement of Trp150 caused the loss of the activity only toward fluoroacetate. An interaction between Trp150 and the fluorine atom is probably an absolute requirement for the reduction of the activation energy for the cleavage of the carbon-fluorine bond. - Atsushi Kurata; Michiyo Fujita; Amr M. Mowafy; Harumi Kamachi; Tatsuo Kurihara; Nobuyoshi EsakiJOURNAL OF BIOSCIENCE AND BIOENGINEERING SOC BIOSCIENCE BIOENGINEERING JAPAN 105 (4) 429 - 431 1389-1723 2008/04 [Refereed]
(S)-2-Chloropropionate is a synthetic intermediate for phenoxypropionic acid herbicides. We constructed a system for asymmetric reduction of 2-chloroacrylate to produce (S)-2-chloropropionate with recombinant Escherichia coli cells producing 2-haloacrylate reductase from Burkholderia sp. WS and an NADPH regeneration system. The system provided 37.4 g/l (S)-2-chloropropionate in more than 99.9% e.e. - Atsushi Kurata; Kohsuke Uchimura; Shigeru Shimamura; Tohru Kobayashi; Koki HorikoshiAPPLIED MICROBIOLOGY AND BIOTECHNOLOGY SPRINGER 77 (2) 311 - 319 0175-7598 2007/11 [Refereed]
The acpI gene encoding an alkaline protease (AcpI) from a deep-sea bacterium, Alkalimonas collagenimarina AC40(T), was shotgun-cloned and sequenced. It had a 1,617-bp open reading frame encoding a protein of 538 amino acids. Based on analysis of the deduced amino acid sequence, AcpI is a subtilisin-like serine protease belonging to subtilase family A. It consists of a prepropeptide, a catalytic domain, and a prepeptidase C-terminal domain like other serine proteases from the genera Pseudomonas, Shewanella, Alteromonas, and Xanthomonas. Heterologous expression of the acpI gene in Escherichia coli cells yielded a 28-kDa recombinant AcpI (rAcpI), suggesting that both the prepropeptide and prepeptidase C-terminal domains were cleaved off to give the mature form. Analysis of N-terminal and C-terminal amino acid sequences of purified rAcpI showed that the mature enzyme would be composed of 273 amino acids. The optimal pH and temperature for the caseinolytic activity of the purified rAcpI were 9.0-9.5 and 45 degrees C in 100 mM glycine-NaOH buffer. Calcium ions slightly enhanced the enzyme activity and stability. The enzyme favorably hydrolyzed gelatin, collagen, and casein. AcpI from A. collagenimarina AC40(T) was also purified from culture broth, and its molecular mass was around 28 kDa, indicating that the cleavage manner of the enzyme is similar to that in E. coli cells. - Atsushi Kurata; Masayuki Miyazaki; Tohru Kobayashi; Yuichi Nogi; Koki HorikoshiINTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY SOC GENERAL MICROBIOLOGY 57 1549 - 1553 1466-5026 2007/07 [Refereed]
A psychrotolerant, obligately alkaliphilic, collagenolytic enzyme-producing bacterium, strain AC40(T), was isolated from a deep-sea sediment off Torishima Island, Japan, at a depth of 4026 m. Phylogenetic analysis of 16S rRNA gene sequences indicated that this bacterium was closely related to members of the genus Alkalimonas, with highest sequence similarity (97.9 %) to Alkalimonas delamerensis 1E1(T). DNA-DNA hybridization experiments of strain AC40(T) with A. delamerensis 1E1(T) revealed a level of relatedness of less than 30%. Cells of strain AC40(T) were strictly aerobic, rod-shaped, Gram-negative and motile by means of a single polar flagellum. The organism grew over a range of temperatures from 5 to 37 degrees C and at initial pH values between 7.0 and 10.5. Optimal growth was observed at 33 degrees C and at pH 8.5-10.0. Cellular fatty acids of strain AC40(T) were predominantly saturated and mono-unsaturated straight-chain components (C-16:0 and C-18:1). The major isoprenoid quinone was Q-8. The G + C content of the DNA was 49.3 mol%. Phylogenetic characteristics, physiological properties and DNA-DNA hybridization data indicate that strain AC40(T) represents a novel species of the genus Alkalimonas, for which the name Alkalimonas collagenimarina sp. nov. is proposed. The type strain is AC40(T) (=JCM 14267(T) =NCIMB 14266(T)). - Tohru Kobayashi; Jie Lu; Zhijun Li; Vo Si Hung; Atsushi Kurata; Yuji Hatada; Ken Takai; Susumu Ito; Koki HorikoshiAPPLIED MICROBIOLOGY AND BIOTECHNOLOGY SPRINGER 75 (1) 71 - 80 0175-7598 2007/05 [Refereed]
A new high-alkaline protease (ALTP) was purified to homogeneity from a culture of the strictly anaerobic and extremely alkaliphilic Alkaliphilus transvaalensis. The molecular mass was 30 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme showed the maximal caseinolytic activity higher than pH 12.6 in KCl-NaOH buffer at 40 degrees C. Hydrolysis of the oxidized insulin B-chain followed by mass spectrometric analysis of the cleaved products revealed that as many as 24 of the total 29 peptide bonds are hydrolyzed in a block-cutting manner, suggesting that ALTP has a widespread proteolytic functions. Calcium ion had no effect on the activity and stability of ALTP, unlike known subtilisins. The deduced amino acid sequence of the enzyme comprised 279 amino acids plus 97 prepropeptide amino acids. The amino acid sequence of mature ALTP was confirmed by capillary liquid chromatography coupled to tandem mass spectrometry, which was the 93% coverage of the deduced amino acid sequence. The mature enzyme showed moderate homology to subtilisin LD1 from the alkaliphilic Bacillus sp. strain KSM-LD1 with 64% identity, and both enzymes formed a new subcluster at an intermediate position among true subtilisins and high-alkaline proteases in a phylogenetic tree of subtilase family A. ALTP is the first high-alkaline protease reported from a strict anaerobe in this family. - A Kurata; T Kurihara; H Kamachi; N EsakiJOURNAL OF BIOLOGICAL CHEMISTRY AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC 280 (21) 20286 - 20291 0021-9258 2005/05 [Refereed]
A soil bacterium, Burkholderia sp. WS, grows on 2-chloroacrylate as the sole carbon source. To identify the enzymes metabolizing 2-chloroacrylate, we carried out comparative two-dimensional gel electrophoresis of the proteins from 2-chloroacrylate- and lactate-grown bacterial cells. As a result, we found that a protein named CAA43 was inducibly synthesized when the cells were grown on 2-chloroacrylate. The CAA43 gene was cloned and shown to encode a protein of 333 amino acid residues (M-r 35,788) that shared a significant sequence similarity with NADPH-dependent quinone oxidoreductase from Escherichia coli (38.2% identity). CAA43 was overproduced in E. coli and purified to homogeneity. The purified protein catalyzed the NADPH-dependent reduction of the carbon-carbon double bond of 2-chloroacrylate to produce (S)-2-chloropropionate, which is probably further metabolized to (R)-lactate by (S)-2-haloacid dehalogenase in Burkholderia sp. WS. NADH did not serve as a reductant. Despite the sequence similarity to quinone oxidoreductases, CAA43 did not act on 1,4-benzoquinone and 1,4-naphthoquinone. 2-Chloroacrylate analogs, such as acrylate and methacrylate, were also inert as the substrates. In contrast, 2-bromoacrylate served as the substrate. Thus, we named this novel enzyme 2-haloacrylate reductase. This study revealed a new pathway for the degradation of unsaturated organohalogen compounds. It is also notable that the enzyme is useful for the production of (S)-2-chloropropionate, which is used for the industrial production of aryloxyphenoxypropionic acid herbicides. - A Kurata; T Kurihara; H Kamachi; N EsakiTETRAHEDRON-ASYMMETRY PERGAMON-ELSEVIER SCIENCE LTD 15 (18) 2837 - 2839 0957-4166 2004/09 [Refereed]
Burkholderia sp. WS has a novel enzyme that catalyzes the asymmetric reduction of 2-chloroacrylic acid to yield (S)-2chloropropionic acid, which is used as a building block for the synthesis of aryloxyphenoxypropionic acid herbicides. NADPH is required as a co-substrate for this reaction. (C) 2004 Elsevier Ltd. All rights reserved. - Macromolecular insect chitinase inhibitors produced by fungi: Screening and partial characterizationT Nitoda; H Usuki; A Kurata; H KanzakiJOURNAL OF PESTICIDE SCIENCE PESTICIDE SCI SOC JAPAN 28 (1) 33 - 36 0385-1559 2003 [Refereed]
Culture broths of fungal strains were screened for novel insect chitinase inhibitors using the Spodoptera litura chitinase inhibitory assay. The culture filtrates of 5 strains showed potent and specific inhibitory activity against the insect chitinase. Partial characterization showed that the active compounds produced by these strains were water-soluble macromolecular compounds which had not been hitherto reported as chitinase inhibitors. These novel chitinase inhibitors are, therefore, expected to be potential agents for insect control.
MISC
- 相松光太; 石田隼斗; 木村友希; 藤井暁; 長野正信; 今井友也; 倉田淳志; 上垣浩一 日本農芸化学会大会講演要旨集(Web) 2022- 2022
- 塩谷瑞紀; 松尾啓史; 大谷里菜; 長谷川哲哉; 大西徹; 井田悠斗; 副田尚位; 定塚勝樹; 倉田淳志; 上垣浩一 日本農芸化学会大会講演要旨集(Web) 2022- 2022
- 松尾啓史; 長谷川哲哉; 笠行康葉; 清水浩美; 大橋正孝; 都築正男; 倉田淳志; 上垣浩一 日本農芸化学会大会講演要旨集(Web) 2021- 2021
- 高木敬信; 笠行康葉; 古家美紀; 大橋正孝; 清水浩美; 倉田淳志; 上垣浩一 日本農芸化学会大会講演要旨集(Web) 2021- 2021
- 松尾啓史; 長谷川哲哉; 塩谷瑞紀; 大谷里菜; 木崎健斗; 中谷未侑; 倉田淳志; 上垣浩一 日本醸造学会大会講演要旨集 2021 (CD-ROM)- 2021
- 長谷川哲哉; 松尾啓史; 高木敬信; 大谷里菜; 塩谷瑞紀; 古家美紀; 大西徹; 木崎健斗; 中谷未侑; 森実優; 倉田淳志; 上垣浩一 日本醸造学会大会講演要旨集 2021 (CD-ROM)- 2021
- 山本和樹; 倉田淳志; 上垣浩一 日本農芸化学会大会講演要旨集(Web) 2021- 2021
- 長谷川哲哉; 大谷里菜; 塩谷瑞紀; 松尾啓史; 大西徹; 古家美紀; 高木敬信; 大橋正孝; 倉田淳志; 上垣浩一 日本生物工学会大会講演要旨集 73rd- 2021
- 福井瑞季; 倉田淳志; 三好柚紀; 山崎思乃; 今井友也; 栗原達夫; 上垣浩一 日本農芸化学会大会講演要旨集(Web) 2019- 2019
- 倉田淳志; 清原昇悟; 宮崎綾乃; 三好柚紀; 山崎思乃; 今井友也; 栗原達夫; 上垣浩一 日本農芸化学会大会講演要旨集(Web) 2019- 2019
- 福井瑞季; 倉田淳志; 三好柚紀; 山崎思乃; 今井友也; 栗原達夫; 上垣浩一 日本生物工学会大会講演要旨集 71st- 2019
- A flavoenzyme from Burkholderia sp. WS that is involved in metabolism of 2-chloroacrylic acidAnoop Vasudevan; 藤田倫代; 倉田淳志; 川本純; 栗原達夫; 江崎信芳 日本生化学会第82回大会 ,京都 2011/09
- 2-クロロアクリル酸資化性細菌 Burkholderia sp. WS における FADH2 依存的水付加酵素 2-ハロアクリル酸ヒドラターゼの機能Anoop Vasudevan; 藤田倫代; 倉田淳志; 川本純; 栗原達夫; 江崎信芳 第28回 日本微量栄養素学会学術集会 ,京都 2011/06
- 炭化水素生産菌を利用したホワイトバイオテクノロジーによる物質生産系の基盤構築倉田 淳志; 岸本 憲明 自然に学ぶものづくり研究助成プログラム最終成果及び会計報告書 2010/11
- KURIHARA Tatsuo; FUJITA Michiyo; MOWAFY Amr M.; KURATA Aatsushi; ESAKI Nobuyoshi 日本生物工学会大会講演要旨集 20- 165 -165 2008
- Alkalimonas collagenimarina AC40T倉田 淳志; 小林 徹; 掘越 2006/10
- 仁戸田 照彦; 臼木 博一; 倉田 淳志 Journal of pesticide science 28- (1) 33 -36 2003
- Kurihara Tatsuo; Kurata Atsushi; Kamachi Harumi; Esaki Nobuyoshi 日本生物工学会大会講演要旨集 14- 96 -96 2002
Books and other publications
- 倉田淳志; 岸本憲明 (Contributorイオン液体と微生物・酵素の利用技術の開発)2019/07 9784781314266
- Deep-Sea: Marine Biology, Geology and Human ImpactAtsushi Kurata; Tohru Kobayashi; Koki Horikoshi (ContributorProtein degradation at deep-sea sediment: collagenolytic enzymes produced from a novel marine bacterium.)2012/10 9781622571734
- Ionic Liquid/Book 2, Biotransformation of underutilized natural resource to valuable compounds in ionic liquid: Enzymatic synthesis of caffeic acid phenethyl ester analogues from immature coffee beans倉田 淳志; 岸本 憲明; 藤田 藤樹夫 (ContributorBiotransformation of Underutilized Natural Resource to Valuable Compounds in Ionic Liquid: Enzymatic Synthesis of Caffeic Acid Phenethyl Ester Analogues from Immature Coffee Beans)Intech, Croatia 2011/07
Lectures, oral presentations, etc.
- Lactiplantibacillus plantarumを用いた細胞外膜小胞の高生産方法の開発前田朝海; 中林遥希; 町村瑞季; 岡野憲司; 川本 純; 河野健一; 上垣浩一; 倉田淳志日本乳酸菌学会 2024 年度大会 2024/07
- 腸内環境の恒常性の維持におけるプロバイオティクス由来の細胞外膜小胞の機能倉田淳志第96回日本生化学会大会 2023/11
- Lachancea thermotoleransの清酒醸造への利用と乳酸生成に関わる乳酸デヒドロゲナーゼの解明中谷 未侑; 大谷 里菜; 大西 徹; 長谷川 哲哉; 片渕 歩真; 北野 未唯; 梅澤 究; 大橋 正孝; 倉田 淳志; 上垣 浩一第96回日本生化学会大会 2023/11
- 等温滴定カロリメトリーを用いた酵母α-イソプロピルリンゴ酸シンターゼとエフェクター分子の結合解析木崎 健斗; 古家 美紀; 北詰 花菜; 大沼 貴之; 倉田 淳志; 上垣 浩一第96回日本生化学会大会 2023/11
- 乳酸菌 RD055328 株由来の GAPDH の特性と細胞外膜小胞の精製藤原 遼; 石崎 真尋; 竹内 慎平; 田村 健人; 倉田 淳志; 山崎 思乃; 今井 友也; 上垣 浩一第96回日本生化学会大会 2023/11
- 乳酸菌細胞外膜小胞の高生産条件の開発前田 朝海; 倉田 淳志; 河野 健一; 上垣 浩一第96回日本生化学会大会 2023/11
- Bifidobacterium dentiumのMVsが示すJurkat細胞の増殖抑制効果前田瑞歩; 入江健太; 岡田美玖; 福田隆志; 川本純; 今井友也; 栗原達夫; 倉田淳志; 上垣 浩一第75回日本生物工学会大会 2023/09
- 乳酸菌が産生する細胞外膜小胞の高生産条件の検討前田朝海; 倉田淳志; 河野健一; 上垣浩一第75回日本生物工学会大会 2023/09
- 腸内環境の恒常性維持における乳酸菌やビフィズス菌が放出する細胞外膜 小胞の役割 [Invited]倉田淳志Cytek ウェビナー 2023/07
- 乳酸菌細胞外膜小胞の高生産条件の検討前田朝海; 倉田淳志; 河野健一; 上垣浩一日本乳酸菌学会 2023/07
- 乳酸菌 RD055328 株由来の GAPDH による TLR2 シグナル伝達経路の活性化藤原 遼; 石崎 真尋; 竹内 慎平; 田村 健人; 倉田 淳志; 山崎 思乃; 今井 友也; 上垣 浩一日本乳酸菌学会 2023/07
- Bifidobacterium dentiumが生産するMVsの特性 [Not invited]前田 瑞歩; 小西 莉子; 入江 健太; 岡田 美玖; 福田 隆志; 川本 純; 今井 友也; 栗原 達夫; 倉田 淳志; 上垣 浩一第74回日本生物工学会大会 2022/10
- 乳酸菌やビフィズス菌由来の新たな発酵産物の特性解明とその応用 [Invited]倉田淳志近畿大学コア研究 健康長寿・未病効果が期待できる新たな 機能性食品の開発を目指した実践研究 令和3年度 研究成果報告会 同時開催:ACTプロジェクト報告会 2022/08
- 乳酸菌が放出する細胞外膜小胞の特性 [Invited]倉田淳志日本乳酸菌学会設立30周年記念シンポジウム 2021/11
- Novel TLR2 ligand of Lactiplantibacillus plantarum BMVs induces the pro/anti-inflammatory cytokines and IgA production [Not invited]Atsushi Kurata2021/11
- 発酵食品や腸管に関連する細菌の細胞外膜小胞の特性 [Invited]倉田淳志日本農芸化学会2021年度大会 2021/03
- 相模湾由来単離株51-CSのメンブランベシクルの特性の解析福井瑞季; 倉田淳志; 三好柚紀; 山崎思乃; 今井友也; 栗原達夫; 上垣浩一第71回日本生物工学会大会 2019/09
- Enzymatic Synthesis of Caffeic Acid Phenethyl Ester Analogues from Underutilized Natural Resource in Ionic Liquid [Not invited]倉田 淳志; 岸本 憲明; 藤田 藤樹夫; 農学部応用生命化学科; 農学部応用生命化学科; UCC 上島珈琲4TH CONGRESS ON IONIC LIQUIDS 2011/06 Hilton Crystal City, Arlington, Virginia, USA 4TH CONGRESS ON IONIC LIQUIDS
A novel consecutive enzymatic conversion was developed for production of caffeic acid phenethyl ester analogues from unused coffee beans. The procedure was comprised of chlorogenate hydrolase and Novozyme435 with [BMIM][NTf2] as the solvent. The analogues exhibited strong antiproliferative activities toward various human tumor cells. - 親水性・疎水性イオン液体存在下で生育する微生物の探索 [Not invited]倉田 淳志; 岸本 憲明; 農学部応用生命化学科2011/03【目的】イオン液体はアニオンとカチオンから構成され、常温常圧で液体の塩である。アニオンとカチオンの組み合わせにより疎水性や親水性を示すイオン液体を調製可能であり、疎水性や親水性を示す化合物を溶解できる。さらに不揮発性や難燃性、熱安定性を示す。イオン液体は優れた反応溶媒であり、イオン液体中で利用可能な酵素の取得が期待される。本研究では親水性・疎水性イオン液体存在下で生育する微生物を探索し、酵素活性の検出を試みた。 【方法・結果】まず天日塩や岩塩、醤油もろみ、キムチ、くさや漬け汁から、耐塩性細菌の獲得を試みた結果、20% NaClを含む培地で生育する6種類の細菌群集を得た。続いてイオン液体を含む培地で生育を検討した結果、くさや漬け汁とキムチ由来の細菌群集はそれぞれ疎水性イオン液体[BMIM][PF6]と親水性イオン液体[EMIM][CF3SO3]を含む培地で生育した。[EMIM][CF3SO3]を含む培養液上清からアルカリフォスファターゼとエステラーゼ活
- イオン液体と水溶液を用いたエチルガレートからオクチルガレート配糖体の2段階酵素合成 [Not invited]倉田 淳志; 岸本 憲明; 農学部応用生命化学科; 農学部応用生命化学科2011/03【目的】 Octyl gallateは高い抗菌、抗ウイルス活性を有するが、不溶性であるため扱いにくい。そこで、Octyl gallateの溶解度を高めるため、Ethyl gallateから2段階の酵素反応でOctyl gallate配糖体を酵素合成する方法を開発した。 【方法および結果】 Octyl gallateを直接配糖化する糖転移酵素を探索したが、見つけることができなかった。そこで、基質をEthyl gallate に変えて配糖化酵素を探索した結果、Sucrose phosphorylaseがEthyl gallate-4-O-α-glucopyranosideに変換することを見出した。得られたEthyl gallate-4-O-α-glucopyranosideと1-Octanolから、イオン液体中でOctyl gallate配糖体に変換できるリパーゼを探索したところ、Lipozyme RMIM反応液から新規ピークを検出した。この生成物を単離し、機器分析からOctyl gallate-4-O-α-glucopyranosideと同定した。供試イオン液体中では、[BMIM][NTf2]が最適(変換率40%)で、親水性イオン液体より疎水性イオン液体中で高い活性を示した。Octyl gallate配糖体はPseudomonas aeruginosaとBacillus subtilis
- イオン液体を用いたCaffeoyl quinic acidからMethyl Dicaffeoyl Quinateへの酵素変換 [Not invited]倉田 淳志; 岸本 憲明; 農学部応用生命化学科; 農学部応用生命化学科; 農学部応用生命化学科; UCC上島珈琲日本農芸化学会2011年度大会 2011/03 日本農芸化学会(京都市、京都女子大学) 日本農芸化学会2011年度大会
【目的】Methyl dicaffeoyl quinate (DCQA-Me) は、プロポリスなどに微量含まれ、DPPHラジカル消去作用、抗がん活性など多様な生理活性をもっている。そこで、コーヒー生豆から抽出した 5-Caffeoyl quinic acid(5-CQA)を出発物質とし、イオン液体(IL)中で二段階の酵素反応でDCQA-Meに変換する方法を開発した。 【方法および結果】まず、5-CQAとメタノールから5-CQA-Meを合成する IL と酵素を探索した。その結果、[Bmim] [NTf2]に5-CQAとメタノールを溶解し、Novozyme 435を40℃で 96 h 反応させると、5-CQA-Meを得ることができた(モル変換率93%)。次に、5-CQA-Meとvinyl caffeateからDCQA-Meに変換できる酵素を探索した結果、Lipozyme TLIM反応液から新規ピークを検出した。単離した生成物は、機器分析から4,5-DCQA-Meと同定した。本酵素は疎水性 IL中ではDCQA-Meを合成したが、親水性ILやt-butyl methyl etherやdioxaneなどの有機溶媒中では合成しなかった。また、疎水性ILに溶解した5-CQA-Meは基質として利用したが、溶解しなかった5-CQA - 揮発性酢酸イソアミルの Escherichia coli K-12 に対する生育阻害機構の解析 [Not invited]倉田 淳志; 岸本 憲明; 農学部応用生命化学科; 農学部応用生命化学科; 農学部応用生命化学科日本農芸化学会2011年度大会 2011/03 日本農芸化学会(京都市、京都女子大学) 日本農芸化学会2011年度大会
【目的】本研究室では市販チーズから単離した酵母 Candida maltosa NP9 が揮発性酢酸イソアミル(IA)を生産し、Aspergillus niger 胞子の発芽を阻害することを報告した1)。本研究では、揮発性 IA の抗菌抗真菌スペクトルを評価するとともに、IA 曝露で生じた E.coli 細胞の構造変化を電子顕微鏡で観察し、2D-DIGEを用いたプロテオーム解析をおこなった。 【方法・結果】vapor-agar contact 法で抗菌スペクトルを試験した結果、IA は供試糸状菌 15 種、酵母 5 種、グラム陽性菌 7 種、陰性菌 2 種全ての生育を阻害した。また、IA は A.niger 胞子の発芽を静菌的に阻害したが、 E.coli と Saccharomyces cerevisiae の生育は殺菌的に阻害した。IA に曝露した菌体を TEM 観察すると、E.coli では細胞内の構造に変化が認められ、S.cerevisiae では細胞内小器官の崩壊が観察された。また、E.coli 抽出タンパク質の 2D-DIGE では、発現の異なるタンパク質スポットが複数確認できた。 1)小俣ら:日本農芸化学会 2010 年度大会講演要 - Isolation of microorganisms growing on media containing hydrophilic or hydrophobic ionic liquid [Not invited]岸本 憲明; 妹尾 文哉; 倉田 淳志イオン液体研究会 2011/01 鳥取市 イオン液体研究会
くさや漬け汁とキムチから10%[EMIM]{CF3SO3]存在下で生育する微生物3株とキムチから5~40%[BMIM][PF6]存在下で生育する微生物5株を分離することに成功した。 - Synthesis of methyl dicaffeoyl quinate with enzymatic transesterification reaction in ionic liquid [Not invited]岸本 憲明; 倉田 淳志; 塩見 成哉; 森田 裕子; 岩井 和也イオン液体研究会 2011/01 鳥取市 イオン液体研究会
5-カフェオイルキナ酸をイオン液体中で酵素を用いてキナ酸のカルボキシ基をメチルエステル化した。得られたカフェオイルキナ酸メチルエステルトカフェ酸ビニルエステルからイオン液体中で酵素を用いて4,5-ジカフェオイルキナ酸メチルエステルを酵素合成することに成功した。 - Two-step enzymatic synthesis of octyl gallate-4-O-α-glucopyranoside from ethyl gallate in ionic liauid and aqueous solution [Not invited]岸本 憲明; 倉田 淳志; 宇野 喜晴; 竹本 慎太郎イオン液体研究会 2011/01 鳥取市 イオン液体研究会
sucrose phosphorylase を用いて ethyl gallate を ethyl gallate-4-O-α-glucopyranosideに変換した。次にイオン液体中でethyl gallate-4-O-α-glucopyranosideと1-octanolからLipozyme RMIMを用いて octyl gallate-4-O-α-glucopyranosideに変換できた。octyl gallate-4-O-α-glucopyranosideをα-glucosidaseで処理すると、octyl gallateが生成し、広い抗菌スぺクトルを示した。 - 揮発性抗真菌物質による食品汚染微生物の制御 [Not invited]安藤 仁; 倉田 淳志; 岸本 憲明; 伊藤 久美子第33回日本分子生物学会年会・第83回日本生化学会大会合同大会 2010/12 神戸市、神戸ポートアイランド 第33回日本分子生物学会年会・第83回日本生化学会大会合同大会
- n-Decanoic acid を水酸化して3-Hydroxydecanoic acid を生産する細菌の探索 [Not invited]冨永 祐希; 倉田 淳志; 岸本 憲明; 斎藤 駿; 本田第62回日本生物工学会大会(2010年) 2010/10 宮崎市、フェニックスシーガイアリゾート 第62回日本生物工学会大会(2010年)
- アルカン資化性酵母Candida maltosa による10-Hydroxy-2(E)-decenoic acid の生産と関連酵素の同定 [Not invited]斎藤 駿; 倉田 淳志; 岸本 憲明; 冨永 祐希; 本田第62回日本生物工学会大会(2010年) 2010/10 宮崎市、フェニックスシーガイアリゾート 第62回日本生物工学会大会(2010年)
- Candida maltosa による10-hydroxy-2(E)-decenoic acid の生産と関連酵素遺伝子のクローニング [Not invited]冨永 祐希; 倉田 淳志; 岸本 憲明; 斎藤 駿; 本田日本農芸化学会2010年度関西支部会 2010/10 奈良市 近畿大学農学部 日本農芸化学会2010年度関西支部会
- 揮発性酢酸イソアミルに曝露したAspergillus niger 胞子の微細構造と抗菌抗真菌スペクトル [Not invited]安藤 仁; 倉田 淳志; 岸本 憲明; 伊藤 久美子2010/10
- 市販チーズから単離したBurkholderia sp. T-34が生産する抗真菌ペプチド [Not invited]原 一浩; 倉田 淳志; 岸本 憲明; 中村 瞳日本農芸化学会2010年度関西支部会 2010/10 奈良市 近畿大学農学部 日本農芸化学会2010年度関西支部会
- イオン液体を用いた3-Cyclohexylpropyl caffeate の連続的酵素合成 [Not invited]竹本 慎太郎; 倉田 淳志; 岸本 憲明; 藤田 藤樹夫; 岩井 和也第14回生体触媒化学シンポジウムin 静岡 2010/09 静岡市 グランシップ静岡 第14回生体触媒化学シンポジウムin 静岡
- イオン液体を用いたCaffeic acid phenethyl ester 類縁体の酵素合成 [Not invited]倉田 淳志; 岸本 憲明; 藤田 藤樹夫; 北村 友紀; 赤井; 嘉明; 竹本; 慎太郎; 岩井 和也第14回生体触媒化学シンポジウムin 静岡 2010/09 静岡市 グランシップ静岡 第14回生体触媒化学シンポジウムin 静岡
- 揮発性酢酸イソアミルに曝露した Aspergillus niger 胞子の微細構造と抗菌抗真菌スペクトル [Not invited]伊藤 久美子; 倉田 淳志; 岸本 憲明; 柴田 有香; 安藤2010/08
- イオン液体存在下で生育する微生物の探索及び酵素活性の検出 [Not invited]妹尾文哉; 倉田 淳志; 岸本 憲明第9回近畿大学環境科学研究会 2010/08 東大阪市 近畿大学薬学部 第9回近畿大学環境科学研究会
- 市販チーズから単離したBurkholderia sp. T-34が生産する抗真菌ペプチド [Not invited]原 一浩; 倉田 淳志; 岸本 憲明; 中村 瞳; 倉田; 淳志; 岸本 憲明第9回近畿大学環境科学研究会 2010/08 東大阪市 近畿大学薬学部 第9回近畿大学環境科学研究会
- アルカン資化性酵母Candida maltosaによる10-hydroxy-2(E)-decenoic acid の生産と関連酵素遺伝子のクローニング [Not invited]冨永 祐希; 倉田 淳志; 岸本 憲明; 斎藤 駿; 本田特殊環境微生物セミナー 2010/07 宇治市 京都大学化学研究所 特殊環境微生物セミナー
- イオン液体を用いた酵素反応の開発 [Not invited]倉田 淳志; 岸本 憲明; 竹本 慎太郎; 尾特殊環境微生物セミナー 2010/07 宇治市 京都大学化学研究所 特殊環境微生物セミナー
- 市販チーズから単離したBurkholderia sp.が生産する抗真菌ペプチド [Not invited]岡本 昌也; 倉田 淳志; 岸本 憲明; 原 一浩; 森山; 亮; 北口 佳栄日本農芸化学会2010年度大会 2010/03 目黒区 東京大学駒場Iキャンパス 日本農芸化学会2010年度大会
- 酢酸イソアミル類縁体による糸状菌胞子発芽阻害活性の評価 [Not invited]小俣 地洋; 倉田 淳志; 岸本 憲明; 岡本 昌也; 畑中; 浩志; 北口 佳栄日本農芸化学会2010年度大会 2010/03 目黒区 東京大学駒場Iキャンパス 日本農芸化学会2010年度大会
- 10-hydroxy-2(E)-decenoic acid高生産変異株の取得と変換条件の最適化 [Not invited]宮川 和志; 倉田 淳志; 岸本 憲明; 吉本 一裕; 冨永; 祐希; 信田; 晃佑; 岩崎; 麻美; 川崎; 健児; 倉田; 淳志; 岸本 憲明日本農芸化学会2010年度大会 2010/03 目黒区 東京大学駒場Iキャンパス 日本農芸化学会2010年度大会
- イオン液体を反応溶媒に用いたカフェ酸誘導体の酵素合成と腫瘍細胞増殖阻害活性 [Not invited]北村 友紀; 倉田 淳志; 岸本 憲明; 森山 達哉; 河村 幸雄; 竹本 慎太郎; 赤井; 嘉明; 丸山; 裕平; 西村; 倫栄; 山本; 寛子; 鵜澤 有希; 岩井 和也日本農芸化学会2010年度大会 2010/03 目黒区 東京大学駒場Iキャンパス 日本農芸化学会2010年度大会
- イオン液体を反応溶媒に用いた3-Cyclohexyl caffeateの連続的な酵素合成 [Not invited]竹本 慎太郎; 倉田 淳志; 岸本 憲明; 北村 友紀; 赤井; 嘉明; 樽井 惇; 岩井 和也日本農芸化学会2010年度大会 2010/03 東京都 東京大学駒場Iキャンパス 日本農芸化学会2010年度大会
- 重油資化能の高い微生物群集における炭化水素非資化性細菌の役割 [Not invited]岸本 憲明; 安藤 仁; 西尾 宜峰; 桶谷 孝太郎; 大前 拓也; 倉田 淳志; 藤田 藤樹夫第25回日本微生物生態学会 2009/11 広島 第25回日本微生物生態学会
- 親水性イオン液体を用いた酵素反応の可能性 [Not invited]倉田 淳志; 岸本 憲明第5回広島大学・海洋研究開発機構・近畿大学 合同セミナー 2009/10 西条市 広島大学理学部 第5回広島大学・海洋研究開発機構・近畿大学 合同セミナー
- 深海底泥由来Bacillus sp. F8のコンドロイチン硫酸分解酵素遺伝子の探索 [Not invited]松本 水緒子; 倉田 淳志; 岸本 憲明; 小林 徹第5回広島大学・海洋研究開発機構・近畿大学 合同セミナー 2009/10 西条市 広島大学理学部 第5回広島大学・海洋研究開発機構・近畿大学 合同セミナー
- Shewanella 属細菌による直鎖炭化水素の生産 [Not invited]安藤 仁; 倉田 淳志; 岸本 憲明; 石邨 浩二第5回広島大学・海洋研究開発機構・近畿大学 合同セミナー 2009/10 西条市 広島大学理学部 第5回広島大学・海洋研究開発機構・近畿大学 合同セミナー
- 細菌による直鎖炭化水素の生産 [Not invited]石邨 浩二; 倉田 淳志; 岸本 憲明; 安藤 仁第8回近畿大学環境科学研究会 2009/08 奈良市 近畿大学農学部 第8回近畿大学環境科学研究会
- Burkholderia sp. WS における 2-クロロアクリル酸代謝機構の解析 [Not invited]栗原 達夫; 倉田 淳志; 藤田 倫代; MOWAFY; Amr M; 倉田; 淳志; 江崎; 信芳第60回日本生物工学会大会(2008) 2008/08 仙台市 東北学院大学 第60回日本生物工学会大会(2008)
- 深海底泥由来Alkalimonas collagenimarina AC40Tのコラーゲン分解酵素の特性 [Not invited]倉田 淳志; 内村 康祐; 宮崎; 征行; 島村; 繁; 能木; 裕一; 小林; 徹; 掘越 弘毅日本農芸化学会2008年度大会 2008/03 名古屋市 名城大学天白キャンパス 日本農芸化学会2008年度大会
- 深海底泥から得られた新菌種 Alkalimonas collagenimarina AC40T由来のコラーゲン分解酵素 [Not invited]倉田 淳志; 内村 康祐; 宮崎; 征行; 島村; 繁; 能木; 裕一; 小林; 徹; 掘越 弘毅極限環境微生物学会2007年度大会 2007/11 福岡市 九州大学西新プラザ 極限環境微生物学会2007年度大会
- 2-クロロアクリル酸代謝関連タンパク質の機能解析 [Not invited]藤田 倫代; 倉田 淳志; 栗原 達夫; 江崎; 蒲池 晴美日本農芸化学会2007年度大会 2007/03 世田谷区 東京農業大学世田谷キャンパス 日本農芸化学会2007年度大会
- 酵素を用いた不斉還元反応による(S)-2-クロロプロピオン酸の生産 [Not invited]藤田 倫代; 倉田 淳志; 栗原 達夫; 江崎; 蒲池 晴美日本農芸化学会2006年度大会 2006/03 京都市 京都女子大学 日本農芸化学会2006年度大会
- Pseudomonas sp. YL の2-クロロアクリル酸代謝に関与する酵素群の同定 [Not invited]上村 忠嗣; 倉田 淳志; 栗原 達夫; 江崎; 蒲池 晴美日本農芸化学会2005年度大会 2005/03 札幌市 札幌コンベンションセンター 日本農芸化学会2005年度大会
- Construction of a bioconversion system for the production of (S)-2-chloropropionate from 2-chloroacrylate with 2-chloroacrylate reductase [Not invited]倉田 淳志; 栗原 達夫; 江崎; 蒲池 晴美第77回日本生化学会大会 2004/10 横浜市 パシフィコ横浜 第77回日本生化学会大会
- 2-クロロアクリル酸不斉還元酵素の諸性質 [Not invited]倉田 淳志; 栗原 達夫; 江崎; 蒲池 晴美日本農芸化学会2004年度大会 2004/03 東広島市 広島大学東広島キャンパス 日本農芸化学会2004年度大会
- Identification of the enzyme catalyzing asymmetric reduction of 2-chloroacrylate from Burkholderia sp. WS [Not invited]倉田 淳志; 栗原 達夫; 江崎; 蒲池 晴美第76回日本生化学会大会 2003/10 横浜市 パシフィコ横浜 第76回日本生化学会大会
- 2-クロロアクリル酸不斉還元酵素の高生産系構築と機能解析 [Not invited]倉田 淳志; 栗原 達夫; 江崎; 蒲池 晴美日本農芸化学会2003年度 関西・中部合同支部大会 2003/10 京都市 京都大学 日本農芸化学会2003年度 関西・中部合同支部大会
- A novel enzyme catalyzing the asymmetric reduction of a carbon-carbon double bond [Not invited]倉田 淳志; 栗原 達夫; 江崎; 蒲池 晴美Japanese-German Biochemistry Meeting 2003 2003/09 SORAT-hotel, Marburg, Germany Japanese-German Biochemistry Meeting 2003
- 炭素-炭素二重結合の不斉還元を触媒する新規酵素の構造と機能 [Not invited]倉田 淳志; 栗原 達夫; 江崎; 蒲池 晴美日本農芸化学会2003年度大会 2003/04 藤沢市 日本大学湘南キャンパス 日本農芸化学会2003年度大会
- Burkholderia sp. WS の 2-クロロアクリル酸代謝に関与するタンパク質の同定 [Not invited]倉田 淳志; 栗原 達夫; 江崎; 蒲池 晴美第75回日本生化学会大会 2002/10 京都市 国立京都国際会館 第75回日本生化学会大会
- 2-クロロアクリル酸の不斉還元を触媒する新規酵素の同定 [Not invited]倉田 淳志; 栗原 達夫; 江崎; 蒲池 晴美第54回日本生物工学会大会(2002) 2002/10 大阪市 グランキューブ大阪(大阪国際会議場) 第54回日本生物工学会大会(2002)
- 炭素-炭素二重結合の不斉還元を触媒する新規酵素の開発 [Not invited]倉田 淳志; 栗原 達夫; 江崎; 芳; 蒲池 晴美日本農芸化学会2002年度 関西支部大会 2002/10 奈良市 近畿大学農学部 日本農芸化学会2002年度 関西支部大会
- Burkholderia sp. WS の 2-クロロアクリル酸代謝経路 [Not invited]倉田 淳志; 倉田 淳志; 栗原; 達夫; 江崎 信芳日本農芸化学会2002年度大会 2002/03 仙台市 東北学院大学 日本農芸化学会2002年度大会
- Identification of proteins involved in the metabolism of 2-chloroacrylate in Burkholderia sp. WS [Not invited]倉田 淳志; 栗原 達夫; 江崎; 芳; 蒲池 晴美5th International Winter School on Chemical Biology 2002/02 Marburger Haus, Hirschegg, Austria 5th International Winter School on Chemical Biology
- 糸状菌由来昆虫キチナーゼ阻害物質の探索および性状比較 [Not invited]仁戸田 照彦; 倉田 淳志; 臼木 博一; 田; 淳志; 神崎 浩日本農芸化学会2001年度 関西・西日本・中四国支部合同支部大会 2001/10 岡山市 岡山大学 日本農芸化学会2001年度 関西・西日本・中四国支部合同支部大会
- 糸状菌の生産する昆虫キチナーゼ阻害物質 [Not invited]仁戸田 照彦; 倉田 淳志; 臼木 博一; 倉田; 淳志; 車谷; 治樹; 神崎 浩日本農芸化学会2001年度大会 2001/03 京都市 立命館大学衣笠キャンパス 日本農芸化学会2001年度大会
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Research Themes
- Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)Date (from‐to) : 2021/04 -2024/03Author : 倉田 淳志細菌が細胞外に放出する膜小胞は、生体の免疫系を賦活化できるため、ワクチンアジュバントへの応用が期待される。しかし膜小胞でどのような免疫応答を増強できるのか不明であり、膜小胞による生体への作用機序も不明である。膜小胞の生産機構は不明であり、免疫賦活能に優れた膜小胞の高生産株は育種されていない。膜小胞の活用には、これらの点を解明する必要がある。 すでに発酵食品や腸から独自に分離した細菌の膜小胞に、IgAやIgGという抗体の生産量を増加できるアジュバント活性を発見し、膜小胞による免疫細胞への作用機序を解明しつつある。そこで新たなアジュバント開発を目的として、独自の膜小胞を対象に①抗体の産生誘導活性を担う物質の同定、②膜小胞に対する腸管細胞層の応答や透過性の解明、③細菌による膜小胞の形成・分泌機構の解明を行う。 R3年度には、乳酸菌やビフィズス菌を対象に、膜小胞中の抗体産生誘導能を示す物質の同定を試みた。具体的には、(1)Lactiplantibacillus属細菌、Bifidobacterium属細菌の膜小胞による、炎症性サイトカインや抗炎症性サイトカインの産生誘導能、抗体IgAの産生誘導能を検出できた。(2)Lactiplantibacillus属細菌の膜小胞中のタンパク質を網羅的に同定し、リポタンパク質Lp19180を見いだした。(3)リポタンパク質Lp19180は、異種発現系で発現させて、各種カラムワークで精製した。精製したリポタンパク質Lp19180を用いて、Toll様受容体2を介したNF-κBの活性化、サイトカインの生産誘導能を確認できた。また、Lactiplantibacillus属細菌、Bifidobacterium属細菌の膜小胞中のタンパク質に注目して、抗体を作成して、膜小胞の検出系の構築を試みた。
- Japan Society for the Promotion of Science:Grants-in-Aid for Scientific ResearchDate (from‐to) : 2014/04 -2018/03Author : Nagao Toshihiro; KISHIMOTO Noriaki; KURATA AtsushiSkin microbiota is concerned in several diseases and healthy condition, we thus focus on Staphylococcus aureus and S. epidermidis. Aeromonas hydrophila converted 9cis-C18:1 to 7cis-C16:1 and 5cis-C14:1. 7cis-C16:1 showed selective anti-bacterial activity which suppressed growth of S. aureus and did not suppress growth of S. epidermidis. So, we attempted purification of 7cis-C16:1. Purification of 7cis-C16:1 was not succeeded through enzymatic process, but ODS column was effective for purification of 7cis-C16:1 (86% purity).
Industrial Property Rights
- 特願2011-6528:Synthesis method of quinate diester and qunate diester analog 2011年/01岸本 憲明, 竹本 慎太郎, 塩見 成哉, 森田 裕子, 倉田 淳志, 岩井 和也, 成田 優作疎水性イオン液体に溶解しないカフェオイルキナ酸を、イオン液体中で酵素を用いて疎水性イオン液体に溶解できるカフェオイルキナ酸メチルエステルに変換した。得られたカフェオイルキナ酸メチルエステルとカフェ酸ビニルエステルから、イオン液体中で酵素を用いて4,5-ジカフェオイルキナ酸メチルエステルに変換した。4,5-ジカフェオイルキナ酸メチルエステルは水溶液中で酵素で加水分解され、4,5-ジカフェオイルキナ酸を生成した。
- 特願2007-296564:新規なコラーゲン分解酵素とその利用 2007年/11倉田 淳志, 小林 徹, 掘越