Affiliation |
Faculty of Agriculture Department of Biochemistry and Applied Biosciences |
Title |
Assistant Professor |
External Link |
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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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.
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Alawi A., Kiyoshi K., Matsumoto M., Yamaguchi T., Narita T., Morita T., Suzuki T., Nakajima-Kambe T.
Bioresource Technology Reports 20 2022.12
Language:Japanese Publishing type:Research paper (scientific journal) Publisher:Bioresource Technology Reports
In this study, a promising bacterial strain for the bio-production of 1,3-propandeiol was isolated as a contaminant from a pilot-scale ethanol-producing bioreactor that used glycerol as a substrate. The isolated strain was identified as Citrobacter braakii and named TB-96. As TB-96 showed a fast growth rate, had strong antibiotic resistance, and could assimilate glycerol to produce high concentrations of 1,3-propanediol, it was used for further investigation to promote the biorefining of waste glycerol generated from biodiesel fuel production. First, to increase the 1,3-propanediol productivity of TB-96, the whole genome was sequenced, and the metabolic pathways were investigated. Next, the synthesis pathways of the main by-products—lactate and formate—were disrupted to construct the strain TB-96-dLF. In fed-batch experiments, TB-96-dLF produced up to 60 g/L 1,3-propanediol using only corn steep liquor as a low-cost nitrogen source.
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醸造における木桶の温故知新 Invited
清 啓自
生物工学会誌 99 ( 12 ) 639 - 639 2021.12
Authorship:Corresponding author Language:Japanese Publishing type:Research paper (scientific journal) Publisher:公益社団法人 日本生物工学会
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Tanaka J, Kiyoshi K, Kadokura T, Suzuki KI, Nakayama S
Journal of bioscience and bioengineering 131 ( 4 ) 396 - 404 2021.4
Language:English Publishing type:Research paper (scientific journal) Publisher:Journal of Bioscience and Bioengineering
2,3,5-Triphenyl tetrazolium chloride (TTC) staining is a method to distinguish the mitochondrial activity of cells based on the color: colorless TTC turns red when under reducing conditions. Although the assay reflects the mitochondrial activity of cells, which enzyme(s) in the electron transport system contribute to TTC reduction has been unclear. TTC staining assays using gene disruptants related to the electron transport system in Saccharomyces cerevisiae revealed those disruptants related to electron transport from each electron donor to ubiquinone (red colonies) and disruptants that were related to ubiquinol-cytochrome c oxidoreductase and cytochrome c oxidase (white colonies). In addition, when the enzyme activities of ubiquinol-cytochrome c oxidoreductase and cytochrome c oxidase were measured using TTC as the electron acceptor, only ubiquinol-cytochrome c oxidoreductase showed TTC reduction activity, and the activity was enhanced by potassium cyanide, an inhibitor of cytochrome c oxidase. These results indicated that ubiquinol-cytochrome c oxidoreductase is involved in TTC reduction in S. cerevisiae. The fermentation profiles of BY4741UΔcor1 and BY4741UΔcox4, which exhibited no TTC staining activity, were almost identical to that of the parental strain BY4741U. However, cell growth and ethanol and succinate production of the ura3-mutated strain BY4741, which also exhibited no TTC staining activity, was altered compared to those of BY4741U, indicating that the fermentation profile varies among strains that show no TTC staining activity. The relationship between uracil metabolism and TTC staining activity was also determined based on metabolome analysis.
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Tada S, Itoh Y, Kiyoshi K, Yoshida N
Journal of bioscience and bioengineering 131 ( 5 ) 509 - 517 2021.1
Language:English Publishing type:Research paper (scientific journal) Publisher:Journal of Bioscience and Bioengineering
Ammonia gas-tolerant extremophilic bacteria capable of growing in atmospheres containing up to 4000 ppm of gaseous ammonia were isolated. These bacteria were capable of growing in nutrient broth containing high concentrations of ammonia water, with growth in medium augmented with 0.1–0.2% ammonia exceeding that in medium without ammonia. The minimal inhibitory concentration of ammonia in the medium was 0.5%. The isolated ammonia gas-tolerant bacterium was moderately alkaliphilic, with optimum growth was observed at pH 9. DNA sequence analysis of the 16SrRNA gene revealed that the isolated bacterium was Bacillus lentus. Furthermore, extremophilic bacteria cultured in a 1300 ppm ammonia gas atmosphere on agar medium containing no nitrogen sources were observed to use ammonia gas for growth. These bacteria were identified as Paenibacillus lentus and Bacillus altitudinis based on 16SrRNA gene sequence analysis. The deodorizing effect of ammonia odor by the isolated bacteria immobilized on sawdust was evaluated. The findings showed that forcing ammonia gas through a column containing B. altitudinis immobilized on sawdust reduced the concentration of ammonia gas by 30% compared to columns containing sawdust only. The isolated bacteria immobilized on wood sawdust lost the capacity deodorization after drying, but this function could be restored with increased moisture. The ammonia gas-tolerant extremophilic bacteria immobilized on sawdust show considerable potential for use in ameliorating malodors associated with outdoor heat-treated toilets.
Presentations 【 display / non-display 】
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Effect of regulator gene and environmental stimulation on Clostridium saccharoperbutylacetonicum strain N1-4 spore porphology.
Chaophaya Panjuy, Kuzahiko Furuya, Naoto Yoshida, Shunichi Nakayama, Keiji Kiyoshi
第28回日本生物工学会九州支部佐賀大会 2022.12.3
Event date: 2022.12.3
Language:English Presentation type:Oral presentation (general)
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Citrobacter braakii TB-96の代謝経路改変による電気発酵における発酵産物の変化
柳瀬卓馬、井上謙吾、吉田ナオト、清 啓自
第74回日本生物工学大会 2022.10.18
Event date: 2022.10.17 - 2022.10.19
Language:Japanese Presentation type:Oral presentation (general)
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アンモニアガス耐性細菌Paenibacillus lentus NH33のアンモニア耐性機構の解明
安東 剛、清 啓自、吉田ナオト
第74回日本生物工学大会 2022.10.18
Event date: 2022.10.17 - 2022.10.19
Language:Japanese Presentation type:Oral presentation (general)
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Growth property of gaseous ammonia-tolerant bacterium, Paenibacillus lentus NH33
○Takeshi ANDO, Panjuy Chaophaya, Keiji KIYOSHI, Naoto YOSHIDA
2022.3.16
Event date: 2022.3.15 - 2022.3.18
Language:Japanese Presentation type:Oral presentation (general)
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Construction of 1,3-propanediol fermentation system from glycerol using microbial electrosynthesis
○Takuma YANASE, Kengo INOUE, Naoto YOSHIDA, Keiji KIYOSHI
2022.3.17
Event date: 2022.3.15 - 2022.3.18
Language:Japanese Presentation type:Oral presentation (general)
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 Clostridium属細菌におけるブタノール発酵と芽胞形成
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
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)