Cloning and Expression of GL-7-ACA Acylase in E.coli / 微生物学通报

ABSTRACT

To facilitate the expression of GL 7 ACA acylase gene in a recombinant E coli , a fragment of the gene, in which the signal peptide was deleted by PCR method, was inserted into a prokaryotic expression vector, pET 28a By colony PCR method screening, a recombinant plasmid pET ACY was obtained and then transformed into the expression host BL21 (DE3) The influences of induction conditions such as IPTG concentration, the time of induction and the induction temperature on the expression of the recombinant protein were investigated Under optimal condition, the enzyme activity could reach 266 U/L Finally, the recombinant GL 7 ACA acylase can be easily isolated to a purity of about 80% by a simple anion ion exchange chromatography with enzyme activity recovery of 50%