文献类型：期刊文献

英文题名：Norditerpenoids biosynthesized by variediene synthase-associated P450 machinery along with modifications by the host cell Aspergillus oryzae

作者：Jiang, Lan[1];Lv, Kangjie[1];Zhu, Guoliang[1];Lin, Zhi[2];Zhang, Xue[1];Xing, Cuiping[1];Yang, Huanting[1];Zhang, Weiyan[1];Wang, Zhixin[1];Liu, Chengwei[3];Qu, Xudong[2];Hsiang, Tom[4];Zhang, Lixin[1];Liu, Xueting[1]

机构：[1]East China Univ Sci Technol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[2]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, State Key Lab Microbial Metab, Shanghai 200240, Peoples R China;[3]Northeast Forestry Univ, Coll Life Sci, Key Lab Saline alkali Vegetat Ecol Restorat, Minist Educ, Harbin, Peoples R China;[4]Univ Guelph, Sch Environm Sci, 50 Stone Rd East, Guelph, ON, Canada

年份：2022

卷号：7

期号：4

起止页码：1142

外文期刊名：SYNTHETIC AND SYSTEMS BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000862501400001)】；

基金：We sincerely acknowledge Prof. Jun-lchi Maruyama at the University of Tokyo for the kind provision of A. oryzae NSAR1, Prof. Katsuya Gomi at Tohoku University provides vectors pUARA2 and pUSA2, and Prof. Yi Tang at the University of California provides S. cerevisiae strains and vector pXW55. We sincerely thank Prof. Hideaki Oikawa at the Hokkaido University and Prof. Pinghua Liu at the Boston University for their valuable suggestions. We gratefully acknowledge the financial support from the National Key Research and Development Program of China (2020YFA0907800 and 2019YFA0906200), the National Natural Science Foundation of China (21907031, 81903529, 21977029, 31720103901, 21877124), the Open Project Funding of the State Key Laboratory of Bioreactor Engineering, the 111 Project (B18022). Genome sequencing and assembly of strain DV17020 were supported by funding from the Natural Science and Engineering Research Council of Canada to Prof. T. Hsiang.

语种：英文

外文关键词：Bifunctional terpene synthases; Norditerpenoids; Cytochrome P450; Aspergillus oryzae; Heterologous expression

摘要：The chemical diversity of terpenoids is typically established by terpene synthase-catalyzed cyclization and diversified by post-tailoring modifications. Fungal bifunctional terpene synthase (BFTS) associated P450 en-zymes have shown significant catalytic potentials through the development of various new terpenoids with different biological activities. This study discovered the BFTS and its related gene cluster from the plant endo-phytic fungus Didymosphaeria variabile 17020. Heterologous expression of the BFTS in Saccharomyces cerevisiae resulted in the characterization of a major product diterpene variediene (1), along with two new minor products neovariediene and neoflexibilene. Further heterologous expression of the BFTS and one cytochrome P450 enzyme VndE (CYP6138B1) in Aspergillus oryzae NSAR1 led to the identification of seven norditerpenoids (19 carbons) with a structurally unique 5/5 bicyclic ring system. Interestingly, in vivo experiments suggested that the cyclized terpene variediene (1) was modified by VndE along with the endogenous enzymes from the host cell A. oryzae through serial chemical conversions, followed by multi-site hydroxylation via A. oryzae endogenous enzymes. Our work revealed that the two-enzymes biosynthetic system and host cell machinery could produce structurally unique terpenoids.