Characterization of a new fusicoccane-type diterpene synthase and an associated P450 enzyme

Submitting author affiliation:
Jinan University, Guangzhou, China

Beilstein Arch. 2022, 202247.

Published 09 Jun 2022

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Fusicoccane-type terpenoids are a subgroup of diterpenoids featured with a unique 5-8-5 ring system. They are widely distributed in nature and possess a variety of biological activities. Up to date, only five fusicoccane-type diterpene synthases have been identified. Here, we identify a two-gene biosynthetic gene cluster containing a new fussicoccane-type diterpene synthase gene tadA and an associated cytochrome P450 gene tadB from Talaromyces wortmannii ATCC 26942. Heterologous expression reveals that TadA catalyzes the formation of a new fusicoccane-type diterpene talaro-7,13-diene. D2O isotope labeling combined with site-directed mutagenesis indicate that TadA might employ a different C2,6 cyclization strategy from the known fussicoccane-type diterpene synthases, in which a neutral intermediate is firstly formed and then protonated by an environmental proton. In addition, we demonstrate that the associated cytochrome P450 enzyme TadB is able to catalyze multiple oxidation of talaro-7,13-diene to yield talaro-6,13-dien-5,8-dione.

Keywords: genome mining; diterpene synthase; gene cluster; site-directed mutagenesis; cytochrome P450 enzyme

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Huang, J.-H.; Lv, J.-M.; Xiao, L.-Y.; Xu, Q.; Lin, F.-l.; Wang, G.-Q.; Chen, G.-D.; Qin, S.-Y.; Hu, D.; Gao, H. Beilstein Arch. 2022, 202247. doi:10.3762/bxiv.2022.47.v1

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