| Project/Area Number |
16H04895
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied microbiology
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| Research Institution | University of Tsukuba |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
吉田 健一 神戸大学, 科学技術イノベーション研究科, 教授 (20230732)
矢嶋 俊介 東京農業大学, 生命科学部, 教授 (90301548)
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| Project Period (FY) |
2016-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000)
Fiscal Year 2019: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2018: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2017: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2016: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000)
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| Keywords | L-glucose / L-gluconate / 枯草菌 / イノシトール / 醗酵生産 / 結晶構造解析 / 部位特異的変異 / L-グルコース / inositol |
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| Outline of Final Research Achievements |
In this study we aimed to construct an L-glucose production system in Bacillus subtilis by heterologous expression of iolM, iolN and lgnH genes to produce L-gluconate from myo-inositol. By using synthetic genes optimized for the codon usages of B. subtilis and a strong promoter, these genes can be expressed in B. subtilis at some extent. We introduced these three genes as an operon on a plasmid to B. subtilis strain deficient for intrinsic inositol catabolic genes, however, conversion of myo-inositol to L-gluconate was not observed. It may be due to low expression of these genes.
On the other hand, to obtain a mutant protein of LgdA (L-glucose dehydrogenase) having reverse reaction activity, we have determined its three-dimensional structure. We identified several residues for substrate binding, of which R178 was proven to be essential for inositol dehydrogenation activity, but not for L-glucose dehydrogenation.
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| Academic Significance and Societal Importance of the Research Achievements |
当初予定していた、枯草菌を宿主としたL-グルコース醗酵生産系の確立には至らなかったが、本研究成果をもとにさらに検討を加えることにより、達成は可能であると考えている。将来的にはmyo-inositolという安価な材料からL-グルコースを生産することが可能になると期待される。
また、新規酵素LgdAの立体構造を明らかにし、その構造・機能相関を解析した点、特にR178残基の変異によりL-グルコースにのみ活性を有する酵素が創出できた点は、学術的に意義がある。
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