Peroxide-Driven Hydroxylation of Small Alkanes Catalyzed by an Artificial P450BM3 Peroxygenase System

被引：51
作者：
Chen, Jie [1 ,2 ,3 ]
Kong, Fanhui [1 ,2 ]
Ma, Nana [1 ,2 ,3 ]
Zhao, Panxia [1 ,2 ,3 ]
Liu, Chuanfei [1 ,2 ]
Wang, Xiling [1 ,2 ]
Cong, Zhiqi [1 ,2 ,3 ]
机构：
[1] Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, CAS Key Lab Biofuels, Qingdao 266101, Shandong, Peoples R China
[2] Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Shandong Prov Key Lab Synthet Biol, Qingdao 266101, Shandong, Peoples R China
[3] Univ Chinese Acad Sci, Beijing 100049, Peoples R China
来源：
ACS CATALYSIS | 2019年 / 9卷 / 08期
基金：
中国国家自然科学基金;
关键词：
alkane hydroxylation; hydrogen peroxide; cytochrome P450; protein engineering; peroxygenase; dual-functional small molecule; CYTOCHROMES P450; SELF-SUFFICIENT; MONOOXYGENASE; OXIDATION; OXIDANTS; METHANE;
D O I：
10.1021/acscatal.9b02507
中图分类号：
O64 [物理化学（理论化学）、化学物理学];
学科分类号：
070304 ; 081704 ;
摘要：
We report the selective hydroxylation of small alkanes with H2O2 catalyzed by an artificial P450 peroxygenase system generated from engineered cytochrome P450BM3 variants in assistance with dual-functional small molecule (DFSM), in which DFSM acts as a general acid-base co-catalyst for activating H2O2. This peroxygenase system exhibited comparable catalytic turnover number (TON) to the fungal peroxygenase AaeUPO, the only known H2O2-dependent natural alkane hydroxylase. Moreover, when compared with evolved/engineered NADPH-dependent P450 variants, the current system yielded similar or even better product formation rates (PFRs) but lower total TONs. The substitution of the highly conserved T268 with amino acids having hydrophobic side chains was identified to play critical roles in improving the hydroxylation activity of the DFSM-facilitated P450BM3 peroxygenase system, which is distinct from NADPH-dependent P450 enzymes. These results offer useful insights into how to tune the catalytic functions and chemistry of P450 peroxygenases.
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收藏
页码：7350 / 7355
页数：11
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