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

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Title
Production of active carboxypeptidase Y of Saccharomyces cerevisiae secreted from methylotrophic yeast Pichia pastoris
Author(s)
H S Ro; Mi Sun Lee; Moon Sun Hahm; Hee Sung Bae; Bong Hyun Chung
Bibliographic Citation
Journal of Microbiology and Biotechnology, vol. 15, no. 1, pp. 202-205
Publication Year
2005
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.
Keyword
carboxypeptidasepichia expressionproteinase Ksecretory
ISSN
1017-7825
Publisher
South Korea
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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