In vitro activation of yeast pro-carboxypeptidase Y expressed as inclusion bodies in Escherichia coli

Abstract

The genes encoding pro-carboxypeptidase Ys (pro-CPYs) from Saccharomyces cerevisiae and Hansenula polymorpha were cloned and expressed in Escherichia coli. Most of the expressed pro-CPY was present in the form of inclusion bodies, accounting for about 35% of the total cellular protein. The inclusion bodies solubilized in 6 M guanidinum chloride were renatured by dilution 1:60 into renaturation buffer. Further refolding and activation were followed by the addition of 60 μg ml−1 proteinase K into the diluted solution. The specific activities of in vitro activated S. cerevisiae and H. polymorpha CPYs were found to be similar, representing about 25% of that of native S. cerevisiae CPY.

DOI: 10.1023/A:1005506119165

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@article{Hahm2004InVA, title={In vitro activation of yeast pro-carboxypeptidase Y expressed as inclusion bodies in Escherichia coli}, author={Moon Sun Hahm and Jung Hoon Bae and Eui Sung Choi and Bong Hyun Chung}, journal={Biotechnology Letters}, year={2004}, volume={21}, pages={881-885} }