成果報告書詳細
管理番号20150000000168
タイトル*平成26年度中間報 バイオマスエネルギー技術研究開発 バイオ燃料製造の有用要素技術開発事業 有用微生物を用いた発酵生産技術の研究開発
公開日2015/6/23
報告書年度2014 - 2014
委託先名日揮株式会社 学校法人君が淵学園崇城大学 一般財団法人バイオインダストリー協会 独立行政法人産業技術総合研究所
プロジェクト番号P13011
部署名新エネルギー部
和文要約
英文要約Title: Bio-Energy Technology Development, Project on the Development of Useful Elemental Technology for Biofuel Production, Development of a fermentation technology using the microorganism (FY2013-FY2016) FY2014 Annual Report

In this project, C5C6 simultaneous saccharification and co-fermentation (SSCF) organism and process will be developed, process design package will be prepared, and economic evaluation will be conducted.

1) Construction of C5C6 co-fermenting microorganisms.
The aim of this study is to construct the efficient ethanologenic yeast and to analyze the genetic information of the strain leading to potent xylose utilization, thermotolerance and fermentation-inhibitor tolerance in the SSCF process. In the FY2014, four thermo-tolerant mutants of Saccharomyces cerevisiae were isolated. Genome analyses of these mutants were performed using the next-generation sequencer (NGS), mutations (SNPs and InDels) in those genomes were comprehensively solved, and we succeeded in identifying one of the causal gene mutations leading to efficient thermotolerance. In addition, several native and mutated xylose isomerases were examined to establish an alternative process for xylose utilization in yeast. One of the xylose isomerase mutants exhibited higher xylose utilization.
Genes of xylose reductase, xylitol dehydrogenase, and xylulose kinase of thermotolerant yeast Kluyveromyces marxianus DMB1 were cloned and used for transformation of the strain to enhance these enzymatic activities. The transformants showed high efficiency of ethanol production.
Screening of the genome library of acid-tolerant yeast Pichia kudriavzevii(syn. Issatchenkia orientalis) in S. cerevisiae gave gene candidates that provide acid-tolerance to S. cerevisiae. Furthermore, various genetically engineered yeast strains were examined by established standardized evaluation methods for fermentation ability. One of the yeast strains showed the highest performance of the ethanol production under the condition.

2) Development of an efficient SSCF process
To achieve the target of FY 2013-2014, ethanol fermentation from NaOH-pretreated sugar cane bagasse in 30L-scale using genetically modified yeast was carried out. Ethanol concentration, yield and productivity were over the target of 4.5%, 85% and 0.6g/l/h, respectively.
To obtain data for high concentration slurry handling and design the pilot scale SSCF system, 2m3 scale slurry handling apparatus was and constructed. Using the apparatus, saccharification, fermentation and simulutaneous saccharification and fermentation were carried out. Design of the pilot scale SSCF system corresponding to use genetically modified yeasts has been completed.
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