title: Cloning and expression of a new rat procarboxypeptidase B gene in Escherichia coli and purification of recombination carboxypeptidase B.
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.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Li
authors.initials: L
authors.lastname: Su-Xia
authors.forename: Zhang
authors.initials: Z
authors.lastname: Yu-Jian
authors.forename: Tian
authors.initials: T
authors.lastname: Li-Ping
authors.forename: Yuan
authors.initials: Y
authors.lastname: Qin-Sheng
authors.forename: Gong
authors.initials: G
authors.lastname: Yi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D043424:Carboxypeptidase B / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000302:isolation & purification; D013379:Substrate Specificity; D014357:Trypsin
chemical_list: D004792:Enzyme Precursors; D011994:Recombinant Proteins; D043424:Carboxypeptidase B; D014357:Trypsin