Cloning and Expression of a New Rat Procarboxypeptidase B Gene in Escherichia Coli and Purification of Recombination Carboxypeptidase B

Title: Cloning and Expression of a New Rat Procarboxypeptidase B Gene in Escherichia Coli and Purification of Recombination Carboxypeptidase B

Volume: 10 Issue: 6

Author(s): Li Su-Xia, ZHANG Yu-Jian, TIAN Li-Ping, YUAN Qin-Sheng and GONG Yi

Affiliation:

Keywords: rat, procarboxypeptidase b, rt-pcr, recombinant carboxypeptidase b, cloning and expression, specific activity, purification, application

Abstract: 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.

Cite this article as:

Su-Xia Li, Yu-Jian ZHANG, Li-Ping TIAN, Qin-Sheng YUAN and Yi GONG, Cloning and Expression of a New Rat Procarboxypeptidase B Gene in Escherichia Coli and Purification of Recombination Carboxypeptidase B, Protein & Peptide Letters 2003; 10 (6) . https://dx.doi.org/10.2174/0929866033478627