Identifying the Minimal Enzymes for Unusual Carbon–Sulfur Bond Formation in Thienodolin Biosynthesis |
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| Authors: | Yaya Wang Jiali Wang Shuqi Yu Fan Wang Dr Hongmin Ma Dr Changwu Yue Minghao Liu Prof?Dr Zixin Deng Prof?Dr Ying Huang Prof?Dr Xudong Qu |
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| Affiliation: | 1. Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, Wuhan University, Wuhan, China;2. State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, 1 Beichen West Road, Beijing, China;3. Guizhou Key Laboratory of Microbial Resources and Drug Development, Zunyi Medical College, 201 Dalian Road, Zunyi, China |
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| 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. |
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| Keywords: | biosynthesis carbon-sulfur bond indole-S-hetero scaffold natural products thienodolin tryptophan amino transferase |
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