文献类型：期刊文献

英文题名：Cloning and expression of a new rat procarboxypeptidase B gene in Escherichia coli and purification of recombination carboxypeptidase B

作者：Li, SX; Zhang, YJ; Tian, LP; Yuan, QS; Gong, Y

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Chinese Acad Sci, Shanghai Inst Biol Sci, Biotechnol Res Ctr, Shanghai 200233, Peoples R China

年份：2003

卷号：10

期号：6

起止页码：581

外文期刊名：PROTEIN AND PEPTIDE LETTERS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000187351500007)】；

语种：英文

外文关键词：rat; procarboxypeptidase B; RT-PCR; recombinant carboxypeptidase B; cloning and expression; specific activity; purification; application

摘要：A new coding sequence of the procarboxypeptidase B gene was obtained from SD rat fresh pancreas by RT-PCR and highly expressed in Escherichia coli in inclusion bodies. The folded procarboxypeptidase B was subjected to trypsin enzymatic cleavage to produce active carboxypeptidase B, subsequently, carboxypeptidase B was effectively purified with anion exchange chromatography DEAE-FF and hydrophobic interaction chromatography Octyl FF, as a result, 40 mg carboxypeptidase B per litre cell culture with specific activity 7.42 u/mg was achieved. Further research showed that the obtained recombinant carboxypeptidase B could substitute carboxypeptidase B isolated from pancreas.