We describe the identification and expression cloning of two novel enzymes, a P-glucanase and an aspartic protease, secreted from the basidiomycetous yeast Phaffia rhodozyma. A cDNA library from P. rhodozyma CBS 6938 was constructed, and full-length cDNA encoding an endo-1,3(4)-beta-glucanase (bg1) and an aspartic protease (pr1) were cloned by expression cloning in Saccharomyces cerevisiae W3124. The bgl cDNA encodes a 424-residue precursor protein with a putative signal peptide. The prl cDNA encodes a 405-residue prepropolypeptide with an 81-residue leader peptide. The aspartic protease was purified and characterized. It has a molecular mass of 36 kDa, an isoelectric point of pH 7.5, a pH activity optimum at 4.0-6.0, and a temperature activity optimum around 40 degrees C. Both enzymes show only low sequence identity to other known enzymes.
|Journal||Applied Microbiology and Biotechnology|
|Publication status||Published - 1999|
Villadsen, I., Bang, ML., & Sandal, T. (1999). Cloning and characterization of an endo-b-1,3(4) glucanase and an aspartic protease from Phaffia rhodozyma CBS 6938. Applied Microbiology and Biotechnology, 51(2), 215-222.