CHARACTERIZATION OF A MYRCENE SYNTHASE FROM SUSPENSION CULTURES OF THE MARINE RED ALGA OCHTODES SECUNDIRAMEA

Abstract

Wise, M. L.1, Rorrer, G. L.2, Polzin, J. J.2, Croteau, R. B.1 1Institute of Biological Chemistry, Washington State University, Pullman, WA. 99164 USA; 2 Department of Chemical Engineering, Oregon State University, Corvallis, OR. 96331USAA monoterpene synthase from suspension cultures of the marine red alga Ochtodes secundiramea is shown to biosynthesize myrcene from geranyl diphosphate (GPP) using cell free extracts. This is the first in vitro characterization of a monoterpene synthase from a marine organism. Myrcene is the likely progenitor of the unusual halogenated monoterpenes characteristic of this marine alga and, as such, represents a key step in the biosynthetic pathway. Based on mechanistic considerations from reaction with the biologically relevant substrate GPP, as well as neryl diphosphate (the cis isomer of GPP) and linalyl diphosphate (LPP), the enzyme appears incapable of catalyzing the isomerization of GPP to LPP, a mechanistic feature of most terrestrial monoterpene synthases, perhaps reflecting its evolutionarily ancient origin. The ability to assay and quantitatively monitor the expression of this enzyme in suspension cultures, under strictly defined growth conditions, presents an unparalleled opportunity to delineate, at the molecular level, factors eliciting the biosynthesis of this class of secondary metabolites, to evaluate the metabolic pathway leading to halogenated monoterpenes and to investigate their role in the chemical ecology of marine algae.