Abstract: Germacrene A synthase(GAS) catalyzes the biosynthesis of germacrene A, which is a key precursor for sesquiterpene lactones. Cloning of a novel full-length cDNA encoding GAS from the medicinal plant Crepidiastrum sonchifolium(designated CsGAS) is reported in this study. The cDNA is 1837 bp long and contains a 1680 bp open reading frame encoding a 559 amino-acid protein. The functional expression of the cDNA in Escherichia coli, as an N-terminal thioredoxin fusion protein, with the pET32a vector yielding a recombinant enzyme. Sequence analysis was used to compare this enzyme with the mechanistically related epi-aristolochene synthase from tobacco, and the effect of possible involvement of a number of amino acids in sesquiterpene synthase on product specificity was also discussed.

Key words: Germacrene A synthase, Germacrene A synthase, Germacrene A synthase, Crepidiastrum sonchifolium, Crepidiastrum sonchifolium, Crepidiastrum sonchifolium, Sesquiterpene synthase, Sesquiterpene synthase, Sesquiterpene synthase, Expression, Expression, Expression, Fusion protein, Fusion protein, Fusion protein