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An Unusual Chimeric Diterpene Synthase from Emericella variecolor and Its Functional Conversion into a Sesterterpene Synthase by Domain Swapping
Author(s) -
Qin Bin,
Matsuda Yudai,
Mori Takahiro,
Okada Masahiro,
Quan Zhiyang,
Mitsuhashi Takaaki,
Wakimoto Toshiyuki,
Abe Ikuro
Publication year - 2016
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201509263
Subject(s) - diterpene , terpene , atp synthase , farnesyl pyrophosphate , isopentenyl pyrophosphate , geranylgeranyl pyrophosphate , terpenoid , stereochemistry , chemistry , heterologous expression , prenyltransferase , pyrophosphate , biosynthesis , biochemistry , enzyme , biology , gene , recombinant dna , prenylation
Di‐ and sesterterpene synthases produce C 20 and C 25 isoprenoid scaffolds from geranylgeranyl pyrophosphate (GGPP) and geranylfarnesyl pyrophosphate (GFPP), respectively. By genome mining of the fungus Emericella variecolor , we identified a multitasking chimeric terpene synthase, EvVS, which has terpene cyclase (TC) and prenyltransferase (PT) domains. Heterologous gene expression in Aspergillus oryzae led to the isolation of variediene ( 1 ), a novel tricyclic diterpene hydrocarbon. Intriguingly, in vitro reaction with the enzyme afforded the new macrocyclic sesterterpene 2 as a minor product from dimethylallyl pyrophosphate (DMAPP) and isopentenyl pyrophosphate (IPP). The TC domain thus produces the diterpene 1 and the sesterterpene 2 from GGPP and GFPP, respectively. Notably, a domain swap of the PT domain of EvVS with that of another chimeric sesterterpene synthase, EvSS, successfully resulted in the production of 2 in vivo as well. Cyclization mechanisms for the production of these two compounds are proposed.