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Title 

Production of active carboxypeptidase Y of Saccharomyces cerevisiae secreted from methylotrophic yeast Pichia pastoris

Authors 

H S RoMi Sun LeeMoon Sun HahmHee Sung BaeBong Hyun Chung

Publisher 

The Korean Society for Applied Microbiology

Issue Date 

2005

Citation 

Journal of Microbiology and Biotechnology, vol. 15, no. 1, pp. 202-205

Keywords 

carboxypeptidasepichia expressionproteinase Ksecretorycarboxypeptidase Ccytochrome P450controlled studyenzyme releaseenzyme synthesisgene expression system

Abstract 

Our previous study showed that the overexpression of carboxypeptidase Y (CPY) of Saccharomyces cerevisiae in Escherichia coli resulted in the formation of insoluble inclusion bodies. To produce soluble CPY, we designed a novel Pichia pastoris expression system in which the following were inserted into expression vectors: three different signal sequences derived from the mating factor a1 of S. cerevisiae, an inulinase of Kluyveromyces marxianus, and the endogenous signal sequence of CPY. The expression vector pHIL-D2-SSinul-proCPY was the most effective in the production of proCPY among the vectors examined. The purified active CPY was obtained from proCPY by treating with proteinase K, followed by Q-Excellose ion-exchange column chromatography.

ISSN 

1017-7825

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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