文献类型：期刊文献

英文题名：Bioamination of alkane with ammonium by an artificially designed multienzyme cascade

作者：Yu, Hui-Lei[1];Li, Tuo[1];Chen, Fei-Fei[1];Luo, Xiao-Jing[1];Li, Aitao[2];Yang, Chao[3,4];Zheng, Gao-Wei[1];Xu, Jian-He[1]

机构：[1]East China Univ Sci & Technol, Lab Biocatalysis & Synthet Biotechnol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Hubei Univ, Coll Life Sci, Hubei Collaborat Innovat Ctr Green Transformat Bi, Hubei Key Lab Ind Biotechnol, 368 Friendship Ave, Wuhan 430062, Hubei, Peoples R China;[3]Nankai Univ, Minist Educ, Key Lab Mol Microbiol & Technol, Tianjin 300071, Peoples R China;[4]Nankai Univ, State Key Lab Med Chem Biol, Tianjin 300071, Peoples R China

年份：2018

卷号：47

起止页码：184

外文期刊名：METABOLIC ENGINEERING

收录：;EI(收录号：20181204933975);WOS:【SCI-EXPANDED(收录号:WOS:000433423600018)】；

基金：This work was financially supported by the National Natural Science Foundation of China (Nos. 21672063 & 21536004), the Fundamental Research Funds for the Central Universities (No. 222201514039), Shanghai Science and Technology Program (No. 15JC1400403), and Open Fund of State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology. We thank Liwen Bianji, Edanz Editing China (www.liwenbianji.cn/ac), for editing the English text of a draft of this manuscript.

语种：英文

外文关键词：Systems biocatalysis; Cascade reaction; C-H amination; Cyclohexylamine

摘要：Biocatalytic C-H amination is one of the most challenging tasks. C-H amination reaction can hardly be driven efficiently by solely one enzyme so far. Thus, enzymatic synergy represents an alternative strategy. Herein, we report an "Artificially Bioamination Pathway" for C-H amination of cyclohexane as a model substrate. Three enzymes, a monooxygenase P450(BM3) mutant, an alcohol dehydrogenase ScCR from Streptomyces coelicolor and an amine dehydrogenase EsLeuDH from Exiguobacterium sibiricum, constituted a clean cascade reaction system with easy product isolation. Two independent cofactor regeneration systems were optimized to avoid interference from the endogenous NADH oxidases in the host E. coli cells. Based on a stepwise pH adjustment and ammonium supplement strategy, and using an in vitro mixture of cell-free extracts of the three enzymes, cyclohexylamine was produced in a titer of 14.9 mM, with a product content of up to 92.5%. Furthermore, designer cells coexpressing the three required enzymes were constructed and their capability of alkane bio-amination was examined. This artificially designed bioamination paves an attractive approach for enzymatic synthesis of amines from accessible and cheap alkanes.