Glucoamylases encoded by variantSaccharomycopsis fibuligera genes: Structure and properties

The genes of two variant glucoamylases (GLA1 and GLU1) ofSaccharomycopsis fibuligera were expressed inSaccharomyces cerevisiae, and biochemical properties of the secreted enzymes were compared. It was found that three amino acid alterations in the signal peptide and N-terminal regions of the variants had no effect on the levels of the secreted enzymes. Amino acid alterations in the C-terminal region of the mature proteins influenced their specific activity, substrate specificity, as well as temperature and pH optima. Because of the glycosylation heterogeneity, the glucoamylases of each gene variant were isolated and purified in two forms (A and B), which were essentially similar in catalytic and physicochemical properties but differed in their thermal stability and ability to renaturate after thermal denaturation.