Isolation of penicillin G acylase from Escherichia coli ATCC 11105 by physical and chemical treatments

✍ Scribed by A Kheirolomoom; M Ardjmand; H Fazelinia; A Zakeri

• Publisher: Elsevier Science
• Year: 2001
• Tongue: English
• Weight: 151 KB
• Volume: 8
• Category: Article
• ISSN: 1369-703X
• DOI: 10.1016/s1369-703x(01)00111-5

✦ Synopsis

Different techniques including toluene-ethanol, guanidine hydrochloride, guanidine hydrochloride-EDTA, lysozyme, lysozyme-EDTA and sonication treatments were used to extract penicillin G acylase from Escherichia coli ATCC 11105 cells. The obtained results show that penicillin G acylase extraction by guanidine-EDTA treatment is more specific in comparison to the other applied methods. The maximum specific activity of the enzyme (5.61 U/mg), i.e. the maximum purification was found when 1 M guanidine + 100 mM EDTA solution was used for penicillin G acylase extraction at culture condition including 0.50% (w/v) yeast extract as carbon source. In such a condition more than 95% of the enzyme was extracted. Increasing yeast extract concentration results in the catabolic repression effect of the carbon source and hence lowers amount of the extracted enzyme. The greater value of the specific activity, i.e. the higher purification of the enzyme, the low cost, non requirement of heating and the simple procedure of guanidine-EDTA treatment, lead us to use this method for penicillin G acylase extraction.