2020 Fiscal Year Final Research Report
Elucidating the molecular mechanisms of adaptive evolution from bacterial nutrient acquisition strategies
Project/Area Number |
17H03794
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Research Category |
Grant-in-Aid for Scientific Research (B)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Applied microbiology
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Research Institution | Nara Institute of Science and Technology |
Principal Investigator |
Yoshida Shosuke 奈良先端科学技術大学院大学, 研究推進機構, 特任准教授 (80610766)
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Project Period (FY) |
2017-04-01 – 2021-03-31
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Keywords | PETase / Ideonella sakaiensis / 遺伝子破壊 |
Outline of Final Research Achievements |
Ideonella sakaiensis is capable of growing on PET as a carbon nutrition. In this study, we explored the mechanism of efficient PET degradation by I. sakaiensis and found things as follows. (1) Mutations were introduced into the substrate binding site of PETase, and it was found that the high PET substrate specificity of PETase is greatly influenced by these amino acid residues. (2) We succeeded in constructing a gene disruption system for I. sakaiensis, and created Δpetase and Δmhetase strains. (3) Proteomic analysis revealed an enzyme with hydrolytic activity against PET oligomers.
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Free Research Field |
応用微生物学
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Academic Significance and Societal Importance of the Research Achievements |
(1)PETaseのPETに対する基質特異性の高さに貢献するアミノ酸残基が明らかとなった。これらを他のPET加水分解酵素に置換導入することで、機能改良につながることが期待できる。(2)I. sakaiensis遺伝子破壊系により、本菌の持つ遺伝子の生理学的な機能解析を行うことが可能となった。また、本菌の代謝系を人工的に改良し、有用物質生産につなげるなど、代謝工学への応用が可能となった。(3)新酵素の発見により、本菌のPET代謝への理解を進めることができるようになった。
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