Identification cloning overexpression and characterization of putative terpene cyclase from Stackebrandtia nassauensis

Abstract

Terpenes are the most extensive and structurally diverse group of natural products useful to human beings, but only a few terpene synthases (TPSs) have been characterised from bacteria so far. Here a soil-dwelling Gram-positive, filamentous actinobacteria, Stackebrandtia nassauensis DSM 44728 germacradienol synthase gene (snas_1127), was chosen based on a BLASTp search with pentalenene newlinesynthase (PS) from Streptomyces exfoliatus UC5319 as a query. The snas_1127 gene of S. nassauensis was cloned by PCR and heterologously expressed in Escherichia coli as an N-terminal-His6-tag protein. Incubation of the recombinant protein, SNAS_1127, with farnesyl diphosphate (FPP) in the presence of Mg2+ gave an exclusive sesquiterpene metabolite, germacradienol; a precursor for many industrially important terpene metabolites, without its further conversion to geosmin or geosmin like hydrocarbons. We report the first evidence of the heterologous expression of the terpene synthase gene from S. nassauensis and the establishment of its biochemical function. newline newline