Acta Chimica Sinica ›› 2020, Vol. 78 ›› Issue (6): 490-503.DOI: 10.6023/A20030086 Previous Articles     Next Articles

Review

细胞色素P450酶催化的烷烃选择性羟化

王曦翎a, 陈杰a,b, 马娜娜a,b, 丛志奇a,b   

  1. a 中国科学院青岛生物能源与过程研究所 中国科学院生物燃料重点实验室 山东省合成生物学重点实验室 青岛 266101;
    b 中国科学院大学 北京 100049
  • 收稿日期:2020-03-24 发布日期:2020-05-08
  • 通讯作者: 丛志奇 E-mail:congzq@qibebt.ac.cn
  • 作者简介:王曦翎,助理研究员,2016年于中国海洋大学医药学院获得医学博士学位,2016年7月至2019年8月在中国科学院青岛生物能源与过程研究所从事博士后研究.2019年9月起于中国科学院青岛生物能源与过程研究所工作.主要从事细胞色素P450酶的生物催化研究;丛志奇,研究员,博士生导师,2009年获得日本熊本大学理学博士学位.2009年至2016年,先后在日本分子科学研究所、名古屋大学从事科学研究工作.2016年05月加入中国科学院青岛生物能源与过程研究所,担任课题组长.2017年入选青岛市创新领军人才.从事金属酶的化学生物学与合成生物学研究,当前主要研究兴趣包括细胞色素P450酶的人工设计与实验室进化、甲烷和二氧化碳生物转化和非天然生物催化反应开发等.主持国家自然科学基金面上项目2项,中国科学院、青岛市和研究所自主部署项目等多项科研课题.
  • 基金资助:
    项目受国家自然科学基金(Nos.21778060,21977104)和青岛市创新领军人才计划(No.18-1-2-9-zhc)资助.

Selective Hydroxylation of Alkanes Catalyzed by Cytochrome P450 Enzymes

Wang Xilinga, Chen Jiea,b, Ma Nanaa,b, Cong Zhiqia,b   

  1. a CAS Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101;
    b University of Chinese Academy of Sciences, Beijing 100049
  • Received:2020-03-24 Published:2020-05-08
  • Supported by:
    Project supported by the National Natural Science Foundation of China (Nos. 21778060, 21977104) and the Qingdao Innovative Leading Talent Project (No. 18-1-2-9-zhc).

The selective oxyfunctionalization of unactivated C-H bonds is one of long-standing issues and current topics in synthetic chemistry. One of the major synthetic targets for these reactions is the direct and selective hydroxylation of alkanes to alcohols, however, which faces many severe challenges in controlling chemoselectivity, regioselectivity and stereoselectivity. In nature, the oxidative metalloenzymes is capable of selectively catalyzing the insertion of oxygen into inert C-H bonds of alkanes, such as methane monooxygenases (MMO), soluble butane monooxygenases (sBMO), fungal peroxygenases and Cytochrome P450 monooxygenases (P450s). Among them, P450s that catalyze a variety of oxygenation reactions have attracted special attentions because of some intrinsic advantages. P450s are widely distributed in plants, animals and microorganisms and over 41000 sequences of P450 genes have been named from various databases, which enhances the potentials of P450s in developing the oxidative biocatalysts. In addition, compared with MMOs, P450s that have smaller molecule weight (≈45 kDa) are simple and amenable to recombinant expression and engineering. Herein, we reviewed the recent progress of alkanes hydroxylation by P450 enzymes either in its natural forms or engineered variants, as well as chemical activated systems. The related background and the catalytic mechanism of P450s for alkanes hydroxylation were firstly discussed. The representative examples by natural P450s mainly from CYP153, CYP52 and other P450 families were then outlined. The strategies of rational design and directed evolution on P450s engineering were then summarized focusing on the native/non-native alkane substrates. Three unusual strategies, including substrate engineering, decoy molecule, and dual-functional small molecule co-catalysis, were also discussed on their applications for activating P450s to hydroxylate non-native small alkanes. Finally, we perspective the challenges and solutions that faced by P450 enzymes in the development of new biocatalytic systems toward selective hydroxylation of alkanes. In conclusion, cytochrome P450 enzymes in both of their native and modified form are promising biocatalysts for alkanes hydroxylation and need further be investigated to gain the practical industrial applications.

Key words: alkane, enzyme catalysis, hydroxylation, cytochrome P450 enzyme, C-H activation, biocatalysis, oxidoreductases, directed evolution