MOLECULAR-CLONING AND EXPRESSION OF BACILLUS-SUBTILIS-BGLS GENE IN SACCHAROMYCES-CEREVISIAE

A 2.7-kb EcoRI DNA fragment carrying a Bacillus subtilis endo-beta-1,3-1,4-glucanase gene (bglS) from the E. coli plasmid pFG1 was cloned into an Escherichia coli/yeast shuttle vector to construct a hybrid plasmid YCSH. The hybrid plasmid was used to transform Saccharomyces cerevisiae, and the bglS gene was expressed. Variation between levels of bglS gene expression in S. cerevisiae was about 2.3-fold, depending on the orientation of the 2.7-kb DNA fragment. Assay of substrate specificity and optimal pH of the enzyme demonstrated that the enzyme encoded by YCSH (bglS) was identical with that found in B. subtilis, but the expression level of bglS gene in S. cerevisiae (YCSH) was much lower than that in E. coli (YCSH).