成果報告書詳細
管理番号20130000000867
タイトル*平成24年度中間年報 バイオマスエネルギー技術研究開発事業 戦略的次世代バイオマスエネルギー利用技術開発事業 (次世代技術開発) 非可食バイオマス由来混合糖からのバイオブタノール生産に関わる基盤技術開発
公開日2015/7/25
報告書年度2012 - 2012
委託先名バイオブタノール製造技術研究組合 国立大学法人東京工業大学
プロジェクト番号P10010
部署名新エネルギー部
和文要約
英文要約Title: Research and Development of Technology for Biomass Energy Conversion / Strategic Development for Next-generation Technology for Utilization of Biomass Energy / Development of Biobutanol Production Fundamental Technology from Non-edible Biomass Resources (FY2010-FY2013) FY2012 Annual Report

[1] Metabolic engineering of Corynebacterium glutamicum for butanol production Promotion of research and development of high biobutanol-producing strain ; 1-1) Search for enzyme with high substrate specificity and high enzymatic activity, 1-2) Conversion of cofactor requirement for adjustment of redox balance, 1-3) Elimination of by-product formation pathways in butanol production, 1-4) Practicality evaluation of a butanol production process. For industrial applications, fermentation of butanol at high temperature offers advantages such as reduction in costs of continuous butanol stripping. We made a trial calculation, when “growth-arrested bioprocess" and “gas-stripping system” were combined, 10 degrees C of reaction temperature was raised, and power energy required for stripping could be reduced by half. Then, we acquired a high temperature mutant which can be grown above 40 degrees C by adaptive evolution. To identify the genes required for tolerance to heat, microarray technology was used to investigate the transcriptome profiling of the thermotorelant mutant in response to heat shock stress. We successfully identified several genes which were differentially expressed in response to heat stress. These data will also be helpful to construct more thermotolerant strains for butanol production in the future. One of the selected strains was found to produce butanol at 42 degrees C better than parental strain at 33 degrees C and at higher productivity, suggesting that the thermotolerant strains is suitable for butanol fermentation at high temperatures. [2] Regulatory mechanism of sugar metabolism by RNase in Corynebacterium glutamicum; We found that the amount of isocitrate lyase, an enzyme of the glyoxylate cycle, increased in NCgl2281 knockout mutant cells grown on sodium acetate as the sole carbon source. The growth of the wild type on minimal medium containing 1% sodium acetate started after a lag phase of 2-3 hours, while such a lag phase was not observed for the NCgl2281 knockout mutant. qRT-PCR analysis showed that the level of aceA mRNA was approximately 3-fold higher in the knockout mutant cells than in the wild type. Such differences were not observed in other mRNAs encoding enzymes involved in acetate metabolism. 3’-RACE analysis suggested that RNase E/G cleaves the aceA mRNA in the 3’-untranslated region (3’-UTR). These findings can be applied for overproduction of enzymes involved in isobutanol production. Overproduction of dihydroxy acid dehydratase involved in isobutanol biosynthesis was performed by using aceA 3’-UTR.
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