Reaction mechanism and regulation of the same-site multistep oxidative cytochrome P450 enzymes involved in natural product biosynthesis

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K16034
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 47050:Environmental and natural pharmaceutical resources-related
• Research Institution: Toho University
• Principal Investigator: Iizaka Yohei 東邦大学, 薬学部, 講師 (40770425)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: シトクロムP450酵素 / 多機能型P450酵素 / 生合成 / マクロライド系抗生物質 / 結晶構造解析

Outline of Final Research Achievements

Multifunctional cytochrome P450 enzymes, which catalyze multistep oxidation reactions, are advantageous for diversifying the structure and bioactivity of compounds because a single enzyme can carry out multiple structural modifications. In this study, we focused on P450 RosC, which iteratively oxidize a single site in a substrate during the biosynthesis of the macrolide. Comparative analysis of the substrate affinity and X-ray crystal structure between the wild type and a mutant that catalyzes only the first oxidation reaction revealed some of the factors that enable and regulate the multistep oxidative modification reaction.

Free Research Field

微生物化学

Academic Significance and Societal Importance of the Research Achievements

野生型RosCと変異体の結晶構造解析により多段階酸化修飾に重要なP450の立体構造を見出すことが出来た。本結果は、多機能型P450を目的の反応だけ触媒するように制御できる可能性を示し、工業的にも重要な多機能型P450を利用した目的物質の選択的且つ効率的な生産に貢献することが期待される。また、、本成果に基づいて基質を1回のみ酸化修飾する大多数のP450を多機能型P450へと機能改変することで、新たな有用物質の創出にも繋がる。