Molecular Design of Biocatalysts for Hydroxylation of Small Alkanes

2016 Fiscal Year Final Research Report

• Project/Area Number: 24225004
• Research Category: Grant-in-Aid for Scientific Research (S)
• Allocation Type: Single-year Grants
• Research Field: Chemistry related to living body
• Research Institution: Nagoya University
• Principal Investigator: Watanabe Yoshihito 名古屋大学, 物質科学国際研究センター, 教授 (10201245)
• Co-Investigator(Renkei-kenkyūsha): SHOJI Osami 名古屋大学, 理学研究科, 准教授 (90379587)
• Project Period (FY): 2012-05-31 – 2017-03-31
• Keywords: ヘム酵素 / シトクロムP450 / ガス状アルカン / ベンゼン / 過酸化水素 / 基質特異性 / 酸化反応

Outline of Final Research Achievements

Cytochrome P450BM3 (P450BM3) isolated from Bacillus megaterium catalyzes the hydroxylation of long-alkyl-chain fatty acids. We have demonstrated that even wild-type P450BM3 can catalyze the hydroxylation of gaseous alkanes such as ethane and propane as well as benzene by using perfluorinated carboxylic acids (PFCs) as decoy molecules.　We also have demonstrated that N-perfluoroacyl amino acids strongly activate wild-type P450BM3 for the hydroxylation of inert alkanes. Furthermore, we showed that substrate-binding-state mimics of hydrogen peroxide-dependent cytochrome P450s prepared by one-point mutagenesis are able to catalyze monooxygenation of non-native substrates. The same mutation was also effective in introducing peroxygenase activity into P450BM3 and P450cam, indicating that a variety of peroxygenases based on P450s can be constructed by one-point mutagenesis.

Free Research Field

生物無機化学