Authors : Kafshnochi Magsoud , Safar Farajnia , Raheb Aboshof and Hanieh Rezaee
Abstract: The PGA gene of Escherichia coli encodes a penicillin G acylase (PGA). Penicillin G acylase 1(PGA, EC 3.5.1.11) is a type II penicillin acylase that hydrolyzes Penicillin G to 6-aminopenicillanic acid (6-APA) and phenyl acetic acid (PAA). PGAs have been found in numerous bacteria and fungi and the PGA of Escherichia coli has been well characterized. In free-living E. coli, PGA is thought to act as a Scavenger enzyme for many different natural esters and amides of PAA and its derivatives, such as hydroxyphenylacetic acid (HPA). In this study E. coli, a member of Enterobacteriasea was investigated for PGA. The main aim of this study was screening of E. coli strains from environmental and clinical specimens for PGA by PCR and then cloning, sequencing and recombinant expression of cloned PGA in the E. coli. Total 280 E. coli isolates were collected from water, soil and clinical specimens. Specimens were transported to the laboratory and then routine tests were done for identification. In this study we found PGA gene in only 6 E. coli isolates. All of PGA positive isolates were from Clinical specimens. The PGA gene from one of the positive isolate were amplified and cloned in pGEM T-easy vector. This clone will be used for further study and production of recombinant Penicillin acylase.
Kafshnochi Magsoud , Safar Farajnia , Raheb Aboshof and Hanieh Rezaee , 2008. Screening and Identification of PGA Producing E. coli Isolates by PCR Technique. Research Journal of Biological Sciences, 3: 1186-1188.