Cloning and functional characterization of three sesquiterpene synthase genes from Chamaecyparis formosensis Matsumura

Terpene synthase (TPS) analysis may contribute to a better understanding of terpenoids biosynthesis and the evolution of phylogenetic taxonomy. Chamaecyparis formosensis Matsumura is an endemic and valuable conifer of Taiwan. Its excellent wood quality, fragrance, and durability make it become the five precious conifers in Taiwan. In this study, three sesquiterpene synthase genes that belong to the TPS-d2 Glade were isolated and characterized through in vitro reaction of recombinant protein and in vivo reaction of Escherichia coli heterologous expression system. The main product of Cf-GerA was germacrene A using GC/MS analysis, while the product of Cf-Aco and Cf-Gor were identified as acora-4(14),8-diene and (5R,6R,10S)-alpha-gorgonene by using NMR analysis. These are the first reported enzymes that biosynthesize acora-4(14),8-diene and (5 R,6 R,10 S)-alpha-gorgonene. Both sesquiterpene synthases may isomerize the farnesyl pyrophosphate substrate to nerolidyl pyrophosphate for further cyclization. Cf-Aco may catalyze 1,6-cyclization of nerolidyl cation while Cf-Gor may catalyze through an uncharged intermediate, isogermacrene A.

Automated summary

Terpene synthase (TPS) analysis was conducted to better understand terpenoid biosynthesis and the evolution of phylogenetic taxonomy. Three sesquiterpene synthase genes were isolated and characterized, with their products identified through GC/MS and NMR analysis. These enzymes biosynthesize acora-4(14),8-diene and (5R,6R,10S)-alpha-gorgonene, and may catalyze further cyclization of the substrate.