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Microbiology and Biotechnology Letters

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Microbiol. Biotechnol. Lett. 1995; 23(2): 150-155

Saccharomyces cerevisiae에서 Absidia zychae 의 Carboxypeptidase A cDNA 의 발현과 특성

Expression of Carboxypeptidase Z cDNA from Absidia zychae in saccharomyces cerevisiae and its characteristics

Abstract

Carboxypeptidase Z(CPZ) cDNA of Absidia zychae was experssed in Saccharomyces cerevisiae. The expressed CPZ(YCPZ) was secreted about 30 mg/l into the medium and has a little higher molecular weight than the wild type CPZ in SDS-PAGE. By the result of N-terminal amino acid sequencing, YCPZ has additional 15 amino acids residues in N-terminus of CPZ. But YCPZ shows no difference with CPZ in enzyme activity and substrate specificity. For the identification of processing mechanism of YCPZ, 36-Arg was changed to 36-Thr by site specific mutagenesis. Mutant YCPZ does not processed at 36-Thr. It was, therefore, concluded that the YCPZ was processed by KEX2. According to endo F treatment, high amount of carbohydrate was N-glycosylated in YCPZ.

Keywords: Carboxypeptidase Z. Saccharomyces cerevisiae, Absidia zychae, KEX2, processing, glycosylation, cDNA, site specific mutagenesis

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