Production and secretion of a biologically active Closterium sex pheromone by Saccharomyces cerevisiae

The cDNA fragment coding for the Closterium sex pheromone (protoplast-release-inducing protein inducer, PR-IP inducer) was inserted into a Saccharomyces cerevisiae–Escherichia coli shuttle vector, under the control of the glucose-repressible alcohol dehydrogenase (ADH2) promoter of yeast. The yeast cells transformed by this construct produced and secreted recombinant PR-IP inducer into the surrounding culture medium. The release of PR-IP from mt+ cells was induced by the recombinant pheromone, whereas amino- and carboxy-terminal truncated forms did not show any biological activity. High levels of asparagine-linked glycosylation in the recombinant pheromone were also confirmed after the treatment by deglycosylation enzymes.