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

被引：46
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.
引用
收藏
页码：7350 / 7355
页数：11
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共 48 条
  • [1] The relationships between cytochromes P450 and H2O2: Production, reaction, and inhibition
    Albertolle, Matthew E.
    Guengerich, F. Peter
    [J]. JOURNAL OF INORGANIC BIOCHEMISTRY, 2018, 186 : 228 - 234
  • [2] Structure of the key species in the enzymatic oxidation of methane to methanol
    Banerjee, Rahul
    Proshlyakov, Yegor
    Lipscomb, John D.
    Proshlyakov, Denis A.
    [J]. NATURE, 2015, 518 (7539) : 431 - 434
  • [3] Modification of the heme active site to increase the peroxidase activity of thermophilic cytochrome P450: A rational approach
    Behera, Rabindra Kumar
    Goyal, Sandeep
    Mazumdar, Shyamalava
    [J]. JOURNAL OF INORGANIC BIOCHEMISTRY, 2010, 104 (11) : 1185 - 1194
  • [4] Engineering cytochrome P450cam into an alkane hydroxylase
    Bell, SG
    Orton, E
    Boyd, H
    Stevenson, JA
    Riddle, A
    Campbell, S
    Wong, LL
    [J]. DALTON TRANSACTIONS, 2003, (11) : 2133 - 2140
  • [5] Cytochromes P450 as promising catalysts for biotechnological application: chances and limitations
    Bernhardt, Rita
    Urlacher, Vlada B.
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2014, 98 (14) : 6185 - 6203
  • [6] Catalytic Asymmetric Oxygenations with the Environmentally Benign Oxidants H2O2 and O2
    Bryliakov, Konstantin P.
    [J]. CHEMICAL REVIEWS, 2017, 117 (17) : 11406 - 11459
  • [7] Utilizing Terminal Oxidants to Achieve P450-Catalyzed Oxidation of Methane
    Chen, Mike M.
    Coelho, Pedro S.
    Arnold, Frances H.
    [J]. ADVANCED SYNTHESIS & CATALYSIS, 2012, 354 (06) : 964 - 968
  • [8] Selective hydroxylation of naphthalene using the H2O2-dependent engineered P450BM3 driven by dual-functional small molecules
    Chen, Zhifeng
    Chen, Jie
    Ma, Nana
    Zho, Haifeng
    Cong, Zhiqi
    [J]. JOURNAL OF PORPHYRINS AND PHTHALOCYANINES, 2018, 22 (9-10) : 831 - 836
  • [9] A self-sufficient peroxide-driven hydroxylation biocatalyst
    Cirino, PC
    Arnold, FH
    [J]. ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2003, 42 (28) : 3299 - 3301
  • [10] Activation of Wild-Type Cytochrorne P450BM3 by the Next Generation of Decoy Molecules: Enhanced Hydroxylation of Gaseous Alkanes and Crystallographic Evidence
    Cong, Zhiqi
    Shoji, Osami
    Kasai, Chie
    Kawakami, Norifumi
    Sugimoto, Hiroshi
    Shiro, Yoshitsugu
    Watanabe, Yoshihito
    [J]. ACS CATALYSIS, 2015, 5 (01): : 150 - 156
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