Regulation of P450 TleB catalytic flow for the synthesis of sulfur-containing indole alkaloids by substrate structure-directed strategy and protein engineering

作     者：Xinying Ge Yan Long Jun Wang Bo Gu Zixuan Yang Yinyin Feng Shuo Zheng Yingying Li Wupeng Yan Heng Song 

作者机构：College of Chemistry and Molecular SciencesWuhan UniversityWuhan 430072China School of Life Sciences and BiotechnologyShanghai Jiao Tong UniversityShanghai 200240China 

出 版 物：《Science China Chemistry》 (中国科学(化学英文版))

年 卷 期：2023年第66卷第11期

页      面：3232-3241页

核心收录：

学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 08[工学] 0805[工学-材料科学与工程（可授工学、理学学位）] 080502[工学-材料学] 0901[农学-作物学] 0703[理学-化学] 0836[工学-生物工程] 10[医学] 

基　　金：supported by the National Key Research and Development Program of China(2018YFA0903200 to H.S.) Wuhan University,Undergraduate Training Programs for Innovation and Entrepreneurship of Wuhan University to Y.L. the Science and Technology Commission of Shanghai the National Natural Science Foundation of China(31900033,21ZR1433700)。 

主　　题：substrate structure-directed strategy P450s coupling reaction thioindole alkaloids 

摘      要：Indole alkaloids have attracted considerable attention from synthetic chemists and biochemists for their structural diversity and important biological activities.Compared with traditional organic synthesis methods,the strategy of using cytochrome P450s extraordinary abilities to selectively activate carbon-hydrogen bonds to assist in the synthesis of various indole alkaloids has the characteristics of short synthetic route,mild conditions and high atomic economy.Here,we utilized P450 monooxygenases HinD and TleB to synthesize a novel 6/5/8 tricyclic product from(S)-N-((S)-1-(4-fluoro-1H-indol-3-yl)-3-hydroxypropan-2-yl)-2-mercapto-3-methylbutanamide through the substrate structure-directed strategy.TleB was more effective in catalyzing C–S coupling,and was used to synthesize a series of 6/5/8 tricyclic indololactam derivatives to provide drug candidates.Interestingly,the S–S coupling product was observed in HinD catalysis,which was a minor product in the wild-type TleB catalysis.With the help of protein engineering,we accurately regulated the catalytic flow and reversed the selectivity of TleB to obtain the S–S coupling product.At the same time,the reaction mechanism was reasonably speculated by means of site blocking and protein-substrate complex analysis.