Novel terpenes generated by heterologous expression of bacterial terpene synthase genes in an engineered Streptomyces host

Abstract

Mining of bacterial genome data has revealed numerous presumptive terpene synthases. Heterologous expression of several putative terpene synthase genes in an engineered Streptomyces host has revealed 13 newly discovered terpenes whose GC-MS and NMR data did not match any known compounds in the spectroscopic databases. Each of the genes encoding the corresponding terpene synthases were silent in their parent microorganisms. Heterologous expression and detailed NMR spectroscopic analysis allowed assignment of the structures of 13 new cyclic terpenes. Among these newly identified compounds, two were found to be linear triquinane sesquiterpenes that have never previously been isolated from bacteria or any other source. The remaining 11 new compounds were shown to be diterpene hydrocarbons and alcohol, including hydropyrene (1), hydropyrenol (2), tsukubadiene (11), and odyverdienes A (12) and B (13) each displaying a novel diterpene skeleton that had not previously been reported.

Computational-guided discovery and characterization of a sesquiterpene synthase from Streptomyces clavuligerus

“…The connectivity of the remaining diastereotropic protons H4, H6, H8, and H9 were readily determined from the long-range 1 H- 13 C coupling constants between the respective protons and their neighboring carbon atoms. Our NMR assignments agree with those recently reported by Yamada et al, except we assigned the resonances at H6 to 0.85 ppm, 1.77 ppm and at H4 to 1.26 ppm, 1.84 ppm (15).…”

“…In the same year, Nakano et al reported the characterization of two additional terpene synthases, a monoterpene cyclase that produced 1,8-cineole and a promiscuous acyclic terpene synthase that produced linalool and nerolidol (13,14). While this manuscript was in the final stages of preparation, the function of an additional terpene synthase, which is a protein variant of the subject of this investigation, was reported to synthesize a linear triquinane of undefined stereochemistry (15). The Significance This paper describes a novel strategy for predicting the function of terpene synthases.…”

“…Terpenoids and molecules containing terpenoid fragments are produced from two basic five-carbon building blocks: isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) (1, 2). IPP and DMAPP are then converted into a series of terpenoid diphosphate esters of increasing chain lengths by chain length selective polyprenyl diphosphate synthases to give geranyl diphosphate (GPP, C 10 ), farnesyl diphosphate (FPP, C 15 ), geranylgeranyl diphosphate (GGPP, C 20 ), and higher five-carbon homologs (3). These molecules are substrates for terpene synthases, which catalyze hydroxylation, elimination, cyclization, and rearrangement reactions to produce much of the structural diversity of terpenoid molecules found in nature (4,5).…”

Terpene synthases are widely distributed in bacteria

“…Compounds produced were evaluated by comparison with the mass spectra of the corresponding reference compounds in the National Institute of Standards and Technology (NIST)/United States Environmental Protection Agency (EPA)/National Institutes of Health (NIH) MS Library (2014 version). The generation of previously unidentified terpene products is described in SI Materials and Methods, whereas the NMR structure elucidation of all previously unidentified compounds is described in another work (54). The large-scale preparation of terpenoid metabolites is described in SI Materials and Methods.…”

Nature as organic chemist

“…My laboratory has published more than a dozen papers in Journal of Antibiotics, covering our studies of both terpenoid and polyketide biosynthesis. [63][64][65][66][67] SABBATICAL AT THE UNIVERSITY OF CAMBRIDGE AND THE JOHN INNES CENTRE In 1989, after 6 years as Chair of the Brown Chemistry Department, I spent a 1-year sabbatical leave at the University of Cambridge. My host was Dr Chris Abell, who had been a postdoc in my laboratory in 1982-1983 before returning to a faculty position in the Department of Chemistry in Cambridge.…”