Induction, Cloning and Functional Expression of a Sesquiterpene Biosynthetic Enzyme, δ-Guaiene Synthase, of Aquilaria microcarpa Cell Cultures

Abstract

A homology-based cloning strategy yielded a cDNA clone presumably encoding δ-guaiene synthase, a sesquiterpene cyclase, from tissue cultures of Aquilaria microcarpa, which were treated with methyl jasmonate. Incubation of cell cultures of the plant with yeast extract also induced transcriptional activation of the sesquiterpene synthase gene. The translated protein of the gene obtained by heterologous expression in Escherichia coli catalyzed the cyclization of farnesyl diphosphate to liberate δ-guaiene with α-guaiene and germacrene A as the minor products. The results obtained in the present study, together with the previously reported results, suggest that two classes of δ-guaiene synthase occur in Aquilaria; the enzyme proteins from A microcarpa and A sinensis liberate germacrene A as a minor product, while the protein from A. crassna generates α-humulene instead of germacrene A.