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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
7
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pubmed:dateCreated |
2006-3-8
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pubmed:abstractText |
A new pro-carboxypeptidase (pCPB) gene was cloned by RT-PCR from SD rat pancreas and its overexpression in Escherichia coli resulted in the formation of inclusion bodies (IBs). The IBs of pCPB were solubilized in 8 M urea and successively refolded by urea gradient gel filtration. Subsequently, the renatured pCPB was digested by trypsin. Recombinant active CPB was obtained by passing through DEAE-FF ion exchange and Sephadex-G100 chromatographic column. Capillary electrophoresis assay showed that the purity of the recombinant CPB (rCPB) exceeded 90%. Further, some properties of rCPB were characterized. The optimum of activity was achieved at pH 7-9. The activity of rCPB was inhibited by typical metal chelating agents (EDTA) and Hg2+, and was activated by Co2+ and heat treatment at 40 degrees C. The two-dimension electrophoresis map of rCPB showed that the pI value of rCPB was 5.35. UV absorbance spectrum of the enzyme showed that an absorbance maximum was at 277 nm.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Carboxypeptidase B,
http://linkedlifedata.com/resource/pubmed/chemical/Cations, Divalent,
http://linkedlifedata.com/resource/pubmed/chemical/Drug Combinations,
http://linkedlifedata.com/resource/pubmed/chemical/Metals, Heavy,
http://linkedlifedata.com/resource/pubmed/chemical/Oils,
http://linkedlifedata.com/resource/pubmed/chemical/P & S Liquid,
http://linkedlifedata.com/resource/pubmed/chemical/Phenols,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Precursors,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Urea
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pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
0929-8665
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
12
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
671-6
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pubmed:meshHeading |
pubmed-meshheading:16522183-Animals,
pubmed-meshheading:16522183-Carboxypeptidase B,
pubmed-meshheading:16522183-Cations, Divalent,
pubmed-meshheading:16522183-Chromatography, Gel,
pubmed-meshheading:16522183-Drug Combinations,
pubmed-meshheading:16522183-Electrophoresis, Gel, Two-Dimensional,
pubmed-meshheading:16522183-Enzyme Stability,
pubmed-meshheading:16522183-Indicator Dilution Techniques,
pubmed-meshheading:16522183-Metals, Heavy,
pubmed-meshheading:16522183-Oils,
pubmed-meshheading:16522183-Phenols,
pubmed-meshheading:16522183-Protein Folding,
pubmed-meshheading:16522183-Protein Precursors,
pubmed-meshheading:16522183-Protein Renaturation,
pubmed-meshheading:16522183-Rats,
pubmed-meshheading:16522183-Recombinant Proteins,
pubmed-meshheading:16522183-Spectrum Analysis,
pubmed-meshheading:16522183-Temperature,
pubmed-meshheading:16522183-Urea
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pubmed:year |
2005
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pubmed:articleTitle |
The renaturation of procarboxypeptidase B by urea gradient gel filtration and some properties of recombinant carboxypeptidase B.
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pubmed:affiliation |
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.
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pubmed:publicationType |
Journal Article
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