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Expression and Purification of Ubiquitin-Specific Protease (UBP1) of Saccharomyces cerevisiae in Recombinant Escherichia Coli  

Na, Kang-In (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Kim, Myoung-Dong (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Min, Won-Ki (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Kim, Jeong-Ah (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Lee, Woo-Jong (Korea Biotechnology Commercialization Center, Korea Institute of Industrial Technology)
Kim, Dae-Ok (Department of Food Science and Technology, Kyung Hee University)
Park, Kyung-Moon (Department of Chemical System Engineering, Hongik University)
Seo, Jin-Ho (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Publication Information
Biotechnology and Bioprocess Engineering:BBE / v.10, no.6, 2005 , pp. 599-602 More about this Journal
Abstract
Truncated form of UBP1, an ubiquitin-specific protease of Saccharomyces cerevisiae, was overexpressed in Escherichia coli. The hexahistidine residue $(His_6)$ was fused to the N-terminus of truncated UBP1 and the corresponding recombinant protein was purified with high yield by immobilized metal affinity chromatography. The truncated form of UBP1 protein was functional to cleave ubiquitinated human growth hormone as substrate. Effects of pH and temperature were investigated in order to optimize deubiquitinating reactions for the truncated UBP1. Optimum temperature and pH for the cleavage reaction were $40^{\circ}C$ and pH 8.0, respectively.
Keywords
ubiquitin-specific protease; UBP1; Saccharomyces cerevisiae; Escherichia coli;
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Times Cited By Web Of Science : 7  (Related Records In Web of Science)
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