Cloning and high expression of glutaryl 7-aminocephalosporanic acid acylase gene from Pseudomonas diminuta.

Kim, Dae-Weon; Yoon, Ki-Hong

The gene coding for the glutaryl 7-aminocephalosporanic acid (GL 7-ACA) acylase from Pseudomonas diminuta KAC-1 was cloned and expressed in Escherichia coli. The acylase gene was composed of 2160 base pairs and encoded a polypeptide of 720 amino acid residues. The E. coli BL21 carrying pET2β, the plasmid construct for high expression of GL 7-ACA acylase gene, produced this enzyme at approx. 30% of the total proteins with 3.2 units activity mg protein-1. Growth at temperature below 31 °C and deletion of signal peptide increased the processing of precursor acylase to active enzyme in the recombinant E. coli cells.

ESCHERICHIA coli; GENETIC engineering; PSEUDOMONAS; AMINO acids; ORGANIC acids; ENZYMES

Biotechnology Letters, 2001, Vol 23, Issue 13, p1067

0141-5492

Academic Journal

10.1023/A:1010554323405