title: Identifying the Minimal Enzymes for Unusual Carbon-Sulfur Bond Formation in Thienodolin Biosynthesis.
abstract: Thienodolin (THN) features a tricyclic indole-S-hetero scaffold that encompasses two unique carbon-sulfur bonds. Although its biosynthetic gene cluster has been recently identified in Streptomyces albogriseolus, the essential enzymes for the formation of C-S bonds have been relatively unexplored. Here, we isolated and characterized a new biosynthetic gene cluster from Streptomyces sp. FXJ1.172. Heterologous expression, systematic gene inactivation, and in vitro biochemical characterization enable us to determine the minimum set of genes for THN synthesis, and an aminotransferase (ThnJ) for catalyzing the downstream conversion of tryptophan chlorination. In addition, we evaluated (and mainly excluded) a previously assumed pivotal intermediate by feeding experiments. With these results, we narrowed down four enzymes (ThnC-F) that are responsible for the two unprecedented C-S bond formations. Our study provides a solid basis for further unraveling of the unique C-S mechanisms.
authors: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, Wuhan University, 185 Donghu Road, Wuhan, 430071, China.
authors: Yaya
authors: Y
authors: Wang
authors: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, Wuhan University, 185 Donghu Road, Wuhan, 430071, China.
authors: Jiali
authors: J
authors: Wang
authors: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, Wuhan University, 185 Donghu Road, Wuhan, 430071, China.
authors: Shuqi
authors: S
authors: Yu
authors: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, Wuhan University, 185 Donghu Road, Wuhan, 430071, China.
authors: Fan
authors: F
authors: Wang
authors: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, Wuhan University, 185 Donghu Road, Wuhan, 430071, China.
authors: Hongmin
authors: H
authors: Ma
authors: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, 1 Beichen West Road, Beijing, 100101, China.|Guizhou Key Laboratory of Microbial Resources and Drug Development, Zunyi Medical College, 201 Dalian Road, Zunyi, 563003, China.
authors: Changwu
authors: C
authors: Yue
authors: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, 1 Beichen West Road, Beijing, 100101, China.
authors: Minghao
authors: M
authors: Liu
authors: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, Wuhan University, 185 Donghu Road, Wuhan, 430071, China.
authors: Zixin
authors: Z
authors: Deng
authors: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, 1 Beichen West Road, Beijing, 100101, China. huangy@im.ac.cn.
authors: Ying
authors: Y
authors: Huang
authors: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, Wuhan University, 185 Donghu Road, Wuhan, 430071, China. quxd@whu.edu.cn.
authors: Xudong
authors: X
authors: Qu
mesh_terms: D002244:Carbon / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D007211:Indoles / Q000737:chemistry / Q000378:metabolism*; D005810:Multigene Family; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics; D013455:Sulfur / Q000737:chemistry / Q000378:metabolism*; D013876:Thiophenes / Q000737:chemistry / Q000378:metabolism*; D000637:Transaminases / Q000235:genetics / Q000378:metabolism
chemical_list: D007211:Indoles; D013876:Thiophenes; C000594502:thienodolin; D013455:Sulfur; D002244:Carbon; C030374:indole; D000637:Transaminases
keywords: biosynthesis; carbon-sulfur bond; indole-S-hetero scaffold; natural products; thienodolin; tryptophan amino transferase