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Affiliation |
Faculty of Agriculture Department of Biochemistry and Applied Biosciences |
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Assistant Professor |
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External Link |
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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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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.
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バイオディーゼル副産物からの1,3-プロパンジオール発酵
清 啓自
生物工学会誌 2020.1
Language：Japanese Publishing type：Research paper (bulletin of university, research institution)
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Murai R., Kiyoshi K., Yoshida N.
Current Microbiology 76 ( 11 ) 1298 - 1305 2019.11
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Current Microbiology
© 2019, Springer Science+Business Media, LLC, part of Springer Nature. Geobacillus thermoglucosidasius NY05 catalyzes calcite single crystal formation at 60 °C by using acetate and calcium. Endospores are embedded at the central part of the calcite single crystal and carbon atoms in the calcite lattice are derived from acetate carbon. Here, we synthesized 21-mer antisense DNA oligonucleotides targeting sporulation transcription factor, acetate-CoA ligase, isocitrate lyase, and malate synthase G mRNAs and evaluated the effect of these oligonucleotides on calcite formation in G. thermoglucosidasius NY05. G. thermoglucosidasius NY05 cells containing antisense DNA oligonucleotides targeting sporulation transcription factor, acetate-CoA ligase, isocitrate lyase, and malate synthase G mRNAs had reduced calcite single crystal formation by 18.7, 50.6, 55.7, and 82.3%, respectively, compared with cells without antisense DNA oligonucleotides. These results support that calcite formation needs endospores as the nucleus to grow, and carbon dioxide generated from acetate, which is metabolized via the glyoxylate pathway and glucogenesis, is supplied to the crystal lattice.
Presentations 【 display / non-display 】
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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)
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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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Effect of wooden barrel for microbiome of rice vinegar fermenation mash.
Taishi Yashima. Kenichiro Ohyama, Naoto Yoshida, Keiji Kiyoshi
2021.10.29
Event date： 2021.10.29 - 2022.10.29
Language：Japanese Presentation type：Oral presentation (general)
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宮崎大学焼酎「薫陶」に至る長い道のり
○清 啓自, 榊原 陽一, 水光 正仁
第73回 日本生物工学会大会 2021.10.28
Event date： 2021.10.27 - 2021.10.29
Language：Japanese Presentation type：Oral presentation (general)
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微生物による蒸気式木材乾燥機廃液の反復式脱色
小森香穂, 鳥居朋香, 亀井一郎, 井上謙吾, ○清啓自
2020年度日本フードファクター学会・日本農芸化学会西日本支部合同大会 (宮崎大学（宮崎）) 2020年度日本フードファクター学会・日本農芸化学会西日本支部合同大会実行委員会
Event date： 2020.11.28 - 2020.11.29
Language：Japanese Presentation type：Oral presentation (general)
Venue：宮崎大学（宮崎）
【目的】 木材の製材加工の乾燥工程において、木材に多量の蒸気を噴射する蒸気式乾燥機が広く利用されているが、この時木材由来の抽出成分を含む暗褐色の廃液（以下木材廃液と略す）が多量に排出される。本研究では、これまでに木材廃液の脱色菌として単離したPenicillium citrinum WD4.1株による木材廃液の脱色処理について報告する。
【方法・結果】 宮崎県内の製材工場において杉を蒸気式乾燥機に供した際の木材廃液に栄養素を追加して廃液培地を調製し、脱色に最適培養条件について検討した。次にジャーファメンターを用い、菌体固定担体を繰り返し用いる反復バッチ式の脱色処理を試験した。本株での脱色には廃液に炭素源と窒素源の添加することで、ジャーファメンターでの最適培養系を構築した。次に初期バッチを滅菌廃液培地で行い、本株を純粋培養系で繁殖させた菌体固定担体の繰り返し利用により、以後は未滅菌でも初期バッチと同等の脱色が可能であった。
Grant-in-Aid for Scientific Research 【 display / non-display 】
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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：21433727 2021.04 - 2023.03
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)