Secretion and glycosylation of Clostridium thermocellum endoglucanase A encoded by the celA gene in Saccharomyces cerevisiae.

The Clostridium thermocellum celA gene encoding endoglucanase A is expressed in Saccharomyces cerevisiae but the enzyme produced from the native celA gene is not secreted. After removal of the bacterial signal peptide-coding sequence, the gene was fused to the promoter and prepro segment of the S. cerevisiae MF alpha 1 gene. This construction directs secretion of active endoglucanase A into the culture medium when introduced in yeast on either replicating or integrating vectors. Secretion of endoglucanase A required growth of transformants on rich medium. The secreted enzyme is a 97,000 Da glycoprotein containing about half of its molecular weight as carbohydrate. This new gene fusion could facilitate further research on protein secretion in yeast by using a cellulase as a marker enzyme.