Terminal hydroxylation by cytochrome P450s induced by dummy substrates

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K05271
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 37010:Bio-related chemistry
• Research Institution: Nagoya University
• Principal Investigator: Ariyasu Shinya 名古屋大学, 理学研究科, 助教 (50586998)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: シトクロムP450 / 擬似基質 / 末端水酸化 / 非天然基質 / CYP153

Outline of Final Research Achievements

In this study, we aimed to develop a method to selectively introduce hydroxyl groups to the primary carbon, which has low chemical reactivity, by adding decoy molecules to cytochrome P450s. Through the screening in propane hydroxylation by several long-chain fatty acid hydroxylase P450s in the presence of various decoy molecules, CYP153A33 can hydroxylate the primary carbon in propane with approximately 80% selectivity to produce 1-propanol. Surprisingly, we found that fluorine-contained decoy molecules, which had not been so effective with P450BM3 originating from Bacillus megaterium, were extremely effective for CYP153A33. This P450 can hydroxylate not only propane, but also butane with high selectivity at primary carbons, and these achievements are expected to be developed into a method for selective hydroxylation of primary carbons of non-native substrates in the future.

Free Research Field

生物無機化学

Academic Significance and Societal Importance of the Research Achievements

本研究で達成したシトクロムP450類（特にCYP153類）でのプロパン等の非天然基質の1級炭素選択的水酸化は、反応性の高い分子内部の2級炭素ではなく、反応性の低い分子末端の1級炭素への反応であり、触媒化学的に希少な化学変換である。これはCYP153類の特徴的な細長い反応空間により、分子末端のみが酵素の反応活性種と接触可能なためであり、このような反応場の有用性は酵素以外の合成触媒系への応用が期待出来る。また、分子の末端官能基化は機能性高分子の原料分子の合成にも有用であり、環境負荷の低い酵素反応を化学工業で利用する礎となる成果と言える。