CARBOXYPEPTIDASE B EXPRESSED BY RECOMBINATION, AND PURIFICATION OF THE SAME

摘要

<P>PROBLEM TO BE SOLVED: To provide a method for producing a protein exhibiting carboxypeptidase B activity from a zymogen. Ž<P>SOLUTION: A vector comprises a nucleotide sequence encoding a pre-protein consisting of a rat pro-carboxypeptidase B fused with a histidine tag and signal peptide at its N-terminal. The protein without substantially containing the carboxypeptidase B pro-peptide, having the carboxypeptidase B activity and lacking C-terminal tyrosine is obtained by a method for transforming a microorganic host organism by the above vector, culturing it, secreting a histidine-tagged pro-carboxypeptidase B in culturing medium, immobilizing the same on a particle metal chelate-compatible matrix bindable with the histidine tag, releasing the protein having the carboxypeptidase B activity into a liquid phase by a trypsin treatment, and purifying the protein having the carboxypeptidase B activity from the liquid phase. Ž<P>COPYRIGHT: (C)2010,JPO&INPIT Ž