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Secretion and glycosylation of Clostridium thermocellum endoglucanase A encoded by the celA gene in Saccharomyces cerevisiae
Author(s) -
Benitez J.,
Silva A.,
Vazquez R.,
Noa M. D.,
Hollenberg C. P.
Publication year - 1989
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320050410
Subject(s) - clostridium thermocellum , biology , cellulase , saccharomyces cerevisiae , signal peptide , gene , yeast , secretion , biochemistry , microbiology and biotechnology , glycoprotein , enzyme , recombinant dna
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 α 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.