Regulation and evaluation of five methanol-inducible promoters in the methylotrophic yeast Candida boidinii

摘要

We isolated the promoter regions of five methanol-inducible genes (P_(AOD1), alcohol oxidase; P_(DAS1), dihydroxyacetone synthase; P_(FDH1), formate dehydrogenase; P_(PMP20), Pmp20; and P_(PMP47), Pmp47) from the candida boidinii genome, and evaluated their strength and studied their regulation using the acid phosphatase gene of Saccharomyces cerevisiae (ScPHO5) as the reporter. Of the five promoters, P_(DAS1) was the strongest methanol-inducible promoter whose strength was approximately 1.5 times higher than that of the commonly used P_(AOD1) in methanol-induced cells. Although the expression of P_(AOD1) and P_(DAS1) was completely repressed by the presence of glucose, formate-induced expression of P_(FDH1) was not repressed by glucose. Expression under P_(PMP47), another methanol-inducible promoter, was highly induced by oleate. The induction kinetics of P_(PMP47) and P_(DAS1) revealed that methanol induces the expression of peroxisome membrane protein Pmp47, earlier than the expression of matrix enzyme dihydroxyacetone synthase (Das1p), and that this information is contained in the promoter region of the respective gene. This is the first report which evaluates several methanol-inducible promoters in parallel in the methylotrophic yeast.