Expression and purification of ubiquitin-specific protease (UBP1) ofSaccharomyces cerevisiae in recombinantEscherichia coli |
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| Authors: | Kang-In Na Myoung-Dong Kim Won-Ki Min Jeong-Ah Kim Woo-Jong Lee Dae-Ok Kim Kyung Moon Park Jin-Ho Seo |
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| Institution: | (1) Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, 151-921 Seoul, Korea;(2) Korea Biotechnology Commercialization Center, Korea Institute of Industrial Technology, 406-130 Incheon, Korea;(3) Department of Food Science and technology, Kyung Hee University, Yongin, 449-701 Kyounggi, Korea;(4) Department of Chemical System Engineering, Hongik University, Jochiwon, 339-701 Choongnam, Korea |
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| Abstract: | Truncated form of UBP1, an ubiquitin-specific protease ofSaccharomyces cerevisiae, was overexpressed inEscherichia coli. The hexahistidine residue (His6) 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°C and pH 8.0, respectively. |
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| Keywords: | ubiquitin-specific protease UBP1 Saccharomyces cerevisiae Escherichia coli |
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