Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Production of Active Carboxypeptidase Y of Saccharomyces cerevisiae Secreted from Methylotrophic Yeast Pichia pastoris

  • RO, HYEON-SU ;
  • LEE, MI-SUN (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • HAHM, MOON-SUN (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • BAE, HEE-SUNG (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • CHUNG, BONG HYUN (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology)
  • Published : 2005.02.01
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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 QExcellose ion-exchange column chromatography.

Keywords

References

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