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Affiliation |
Faculty of Agriculture Region of Applied Biochemistry and Biotechnology |
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Title |
Assistant Professor |
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External Link |
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Related SDGs |
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KIYOSHI Keiji
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Degree 【 display / non-display 】
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醸造学 ( 2016.3 東京農業大学 )
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Fermentation technology ( 2012.3 Tokyo University of Agriculture )
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Fermentation technology ( 2010.3 Tokyo University of Agriculture )
Research Areas 【 display / non-display 】
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Life Science / Applied microbiology
Papers 【 display / non-display 】
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Tofu whey treatment and electricity generation by stacked microbial fuel cells Reviewed
Wachid Mochammad, Soichiro Fujimura, Misaki Shichida, Miyuki Nagamine, Keiji Kiyoshi, Kengo Inoue
Journal of Environmental Chemical Engineering 13 ( 3 ) 2025.3
Language：English Publishing type：Research paper (scientific journal)
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Physiological role of the EHL gene in sake yeast and its effects on quality of sake. Reviewed
Tomonaga K, Tanaka J, Kiyoshi K, Akao T, Watanabe K, Kadokura T, Nakayama S
Journal of bioscience and bioengineering 137 ( 3 ) 195 - 203 2024.3
Language：English Publishing type：Research paper (scientific journal) Publisher：Journal of Bioscience and Bioengineering
The EHL1/2/3 genes were identified by whole-genome sequencing of Kyokai No. 7 (K7), which is a well-known representative Japanese sake yeast Saccharomyces cerevisiae. The genes are present in K7, but not in laboratory strain S288C. Although the genes were presumed to encode epoxide hydrolase based on homology analysis, their effect on cellular metabolism in sake yeast has not yet been clarified. We constructed ehl1/2/3 mutants harboring a stop codon in each gene using the haploid yeast strain H3 as the parental strain, which was derived from K701, and investigated the physiological role and effects of the EHL1/2/3 genes on sake quality. Metabolome analysis and vitamin requirement testing revealed that the EHL1/2/3 genes are partly responsible for the synthesis of pantothenate. For fermentation profiles, ethanol production by the ehl1/2/3 mutant was comparable with that of strain H3, but succinate production was decreased in the ehl1/2/3 mutant compared to strain H3 when cultured in yeast malt (YM) medium containing 10% glucose and during sake brewing. Ethyl hexanoate and isoamyl acetate levels in the ehl1/2/3 mutant strain were decreased compared to those of strain H3 during sake brewing. Thus, the EHL1/2/3 genes did not affect ethanol production but did affect the production of organic acids and aromatic components during sake brewing.
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Investigation of diversity in wooden barrel soy sauce(KIOKE SHOYU) using component analysis and sensory evaluation
Kadonosono Tomoko, Yoshida Shiori, Kiyoshi Keiji, Sakaguchi Naoto, Sugimoto Masahiro
Abstracts of the Annual Meeting of the Japan Society of Cookery Science 35 ( 0 ) 86 2024
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：The Japan Society of Cookery Science
[Objective] The traditional method of fermenting soy sauce in wooden barrels (KIOKE SHOYU) was common until the Edo period. However, wooden barrels(KIOKE) have been largely replaced by FRP and concrete tanks due to cost-effectiveness and the difficulty of stable production from the Meiji era. Recently, KIOKE SHOYU has gained recognition as a high-value product due to its "unique and complex flavors specific to each brewery," though scientific knowledge about its components is limited. This study aims to scientifically investigate the diversity of KIOKE SHOYU as part of KIOKE SHOYU EXPORT FACILITATION CONSORTIUM (referred to as "KIOKE Consortium").[Methods] We analyzed 24 types of KIOKE SHOYU from 24 companies (across 17 prefectures) that are members of KIOKE Consortium and 6 types of tank-fermented soy sauce (referred to as "Tank soy sauce") from 5 major manufacturers. The analysis included aroma components using the ultra-fast GC Heracles II, and free amino acids and organic acids using LC-MS. For six distinctive soy sauces, we conducted analytical sensory evaluations and performed principal component analysis (PCA) using the sensory evaluation data correlated with component analysis values. Additionally, we compared the microbial flora between KIOKE and plastic barrels through soy sauce brewing tests.[Results] The PCA results indicated that KIOKE SHOYU were positioned farther apart compared to Tank soy sauce, suggesting greater "variety" in terms of component content. The flavor characteristics of KIOKE SHOYU included types with a strong sweet and roasted aroma and those with complex flavors. The brewing tests showed differences in microbial flora between KIOKE and plastic barrels. Electron microscope observations confirmed the presence of numerous microorganisms on the surface of KIOKE, suggesting that these microorganisms contribute to the unique flavors specific to each brewery.
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Furuya K, Kiyoshi K, Punjuy C, Yoshida N, Maruyama R, Yasuda T, Watanabe K, Kadokura T, Nakayama S
Journal of bioscience and bioengineering 136 ( 3 ) 198 - 204 2023.9
Language：English Publishing type：Research paper (scientific journal) Publisher：Journal of Bioscience and Bioengineering
Clostridium saccharoperbutylacetonicum strain N1-4 (ATCC13564) is a butanol-producing strain suitable for application to butanol production from cellulosic materials by co-culture with cellulolytic and thermophilic species, such as Hungateiclostridium thermocellum (synonym: Clostridium thermocellum). The optimal temperature for butanol production by strain N1-4 is 30 °C, and the strain is sensitive to a high culture temperature of 37 °C. Given that spore formation is observed at high frequency when strain N1-4 is cultivated at 37 °C, we assumed in a previous study that the initiation of sporulation is related to a decrease in butanol production. Therefore, to investigate the relationship between butanol production and spore formation, we generated strain N1-4 isolates in which genes related to spore formation were disrupted. The sporulation-related gene disruptants of spo0A, sigE, sigG, and sigK lost the ability to produce heat-resistant spores, irrespective of the culture temperature. Among the gene disruptants produced, only the spo0A disruptant lost butanol-producing ability when cultivated at 30 °C. Interestingly, the sigE disruptant maintained butanol productivity similar to that observed at 30 °C, even when cultivated at 37 °C. In addition, the sigE disruptant successfully produced butanol from Avicel cellulose by co-culture with H. thermocellum at a fermentation temperature of 37 °C.
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Takase S, Tomonaga K, Tanaka J, Moriya C, Kiyoshi K, Akao T, Watanabe K, Kadokura T, Nakayama S
Journal of bioscience and bioengineering 136 ( 1 ) 44 - 50 2023.5
Language：English Publishing type：Research paper (scientific journal) Publisher：Journal of Bioscience and Bioengineering
Biotin is an essential coenzyme that is bound to carboxylases and participates in fatty acid synthesis. The fact that sake yeast exhibit biotin prototrophy while almost all other Saccharomyces cerevisiae strains exhibit biotin auxotrophy, implies that biotin prototrophy is an important factor in sake brewing. In this study, we inserted a stop codon into the biotin biosynthetic BIO3 gene (cording for 7,8-diamino-pelargonic acid aminotransferase) of a haploid sake yeast strain using the marker-removable plasmid pAUR135 and investigated the fermentation profile of the resulting bio3 mutant. Ethanol production was not altered when the bio3 mutant was cultured in Yeast Malt (YM) medium containing 10% glucose at 15 °C and 30 °C. Interestingly, ethanol production was also not changed during the sake brewing process. On the other hand, the levels of organic acids in the bio3 mutant were altered after culturing in YM medium and during sake brewing. In addition, ethyl hexanoate and isoamyl acetate levels decreased in the bio3 mutant during sake brewing. Metabolome analysis revealed that the decreased levels of fatty acids in the bio3 mutant were attributed to the decreased levels of ethyl hexanoate. Further, the transcription level of genes related to the synthesis of ethyl hexanoate and isoamyl acetate were significantly reduced. The findings indicated that although the decrease in biotin biosynthesis did not affect ethanol production, it did affect the synthesis of components such as organic acids and aromatic compounds. Biotin biosynthesis ability is thus a key factor in sake brewing.
Presentations 【 display / non-display 】
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1 3 プロパンジオール生産性 Citrobacter braakii TB-96 の育種による代謝改変
木村 武蔵, 鮫島 礼, 柳瀬 卓馬, 吉田 ナオト, 中島（神戸） 敏明, 清 啓自
第30回 日本生物工学会九州支部宮崎大会 2025.12.1
Event date： 2025.12.1
Language：Japanese Presentation type：Oral presentation (general)
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タンパク質を基質としたアンモニア生成菌のスクリーニング法
川原 朋、吉田 ナオト、清 啓自
第30回 日本生物工学会九州支部宮崎大会 2025.12.1
Event date： 2025.12.1
Language：Japanese Presentation type：Oral presentation (general)
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Metabolic engineering of 1,3-propanediol productivity in Citrobacter braakii TB-96 using a high-expression promoter
2025.3.11
Event date： 2025.3.5 - 2025.3.8
Language：Japanese Presentation type：Poster presentation
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Enhanced 1,3-propanediol production by cathodic electro-fermentation with Citrobacter braakii strain TB-96 and microbial electron donation by application of anodic electro-fermentation. International conference
Takuma Yanase, Kengo Inoue, Naoto Yoshida, Toshiaki Nakajima-Kambe, Keiji Kiyoshi
IUMS 2024 2024.10.25
Event date： 2024.10.23 - 2024.10.25
Language：English Presentation type：Poster presentation
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グリセロールからの 1,3-プロパンジオール発酵に最適なプロモーターの選抜
木村 武蔵、鮫島 礼、柳瀬 卓馬、吉田 ナオト、中島(神戸) 敏明、 清 啓自
環境バイオテクノロジー学会 2024 年度大会 2024.5.30
Event date： 2024.5.30 - 2024.5.31
Language：Japanese Presentation type：Poster presentation
Grant-in-Aid for Scientific Research 【 display / non-display 】
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微生物の発酵をコントロールする栄養たち
Grant number：23HT0192 2023.04 - 2024.03
独立行政法人日本学術振興会 科学研究費補助金 ひらめきときめきｻｲｴﾝｽ
Authorship：Principal investigator
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核酸代謝の増強が環境ストレスへの対応に及ぼす影響
Grant number：21K14773 2021.04 - 2024.03
独立行政法人日本学術振興会 科学研究費補助金 若手研究
Authorship：Principal investigator
全ての生物は自身の周辺環境から受けるストレスに適応しながら活動、生存し繁殖していく。環境から受けるストレスとは避けられないものであり、そこへ如何に適応できるのかが結果として生物の
種の強さを決定づけているといえる。これは単細胞生物である微生物においては尚更であり、特に発酵に用いられる微生物では如何にこのストレスによる発酵の低下を避けるかは生産性を左右する極めて重要な課題である。外的ストレスが過剰であると微生物は休眠化あるいは細胞死が生じる。
本研究では発酵性の微生物において申請者がこれまでに発見した培地による核酸供給による
熱ストレス耐性の知見を基に、ストレス環境下における核酸などの主要代謝物の外的な供給および生合成の強化によって、いかなるストレスに対してもストレス耐性を付与することが可能であるかを分子生物学的アプローチによって試みる。 -
バイオ燃料廃棄物系バイオマスからポリマー原料への微生物転換
Grant number：99999999 2017.04 - 2019.03
科学研究費補助金 科学技術振興機構 先端的技術開発
Authorship：Coinvestigator(s)
Available Technology 【 display / non-display 】
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農業廃棄物からのブタノール生産
バイオディーゼル燃料の副産物からのアルコール生産
木質が微生物発酵に及ぼす影響の解明Home Page： 微生物機能開発学研究室：
Related fields where technical consultation is available：酒類や発酵食品の醸造技術、様々な原料の微生物への発酵利用
Message：微生物の発酵技術は、様々な物質生産へと応用できます。食品の産業廃棄物など現在利用されていない資源の有効活用法などで微生物の活用を検討の際にお気軽にご相談ください。(清: kiyoshi.keiji.p8@cc.miyazaki-u.ac.jp)