Jiang Lan, Lv Kangjie, Zhu Guoliang, Lin Zhi, Zhang Xue, Xing Cuiping, Yang Huanting, Zhang Weiyan, Wang Zhixin, Liu Chengwei, Qu Xudong, Hsiang Tom, Zhang Lixin, Liu Xueting
State Key Laboratory of Bioreactor Engineering, East China University of Science of Technology, Shanghai, China.
State Key Laboratory of Microbial Metabolism and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China.
Synth Syst Biotechnol. 2022 Aug 23;7(4):1142-1147. doi: 10.1016/j.synbio.2022.08.002. eCollection 2022 Dec.
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 enzymes 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 endophytic fungus 17020. Heterologous expression of the BFTS in resulted in the characterization of a major product diterpene variediene (), along with two new minor products neovariediene and neoflexibilene. Further heterologous expression of the BFTS and one cytochrome P450 enzyme VndE (CYP6138B1) in NSAR1 led to the identification of seven norditerpenoids (19 carbons) with a structurally unique 5/5 bicyclic ring system. Interestingly, experiments suggested that the cyclized terpene variediene () was modified by VndE along with the endogenous enzymes from the host cell through serial chemical conversions, followed by multi-site hydroxylation via endogenous enzymes. Our work revealed that the two-enzymes biosynthetic system and host cell machinery could produce structurally unique terpenoids.
萜类化合物的化学多样性通常由萜烯合酶催化的环化作用建立，并通过后期修饰作用得以多样化。与真菌双功能萜烯合酶（BFTS）相关的P450酶通过开发具有不同生物活性的各种新萜类化合物，已显示出显著的催化潜力。本研究从植物内生真菌17020中发现了BFTS及其相关基因簇。BFTS在[具体宿主]中的异源表达导致鉴定出一种主要产物二萜类化合物variediene（[具体结构]），以及两种新的次要产物新variediene和新flexibilene。BFTS和一种细胞色素P450酶VndE（CYP6138B1）在[具体宿主]NSAR1中的进一步异源表达导致鉴定出七种具有独特结构的5/5双环系统的降二萜类化合物（19个碳原子）。有趣的是，[相关]实验表明，环化萜类化合物variediene（[具体结构]）被VndE以及宿主细胞[具体宿主]NSAR1的内源酶通过一系列化学转化进行修饰，随后通过[具体宿主]NSAR1的内源酶进行多位点羟基化。我们的工作表明，双酶生物合成系统和宿主细胞机制可以产生结构独特的萜类化合物。
