Development and characterization of a potent producer of penicillin G amidase by mutagenization |
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Authors: | S. Quratulain B. Nasira M.A. Kashmiri |
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Affiliation: | (1) Biotechnology and Food Research Center, Pakistan Council of Scientific and Industrial, Research Laboratories (PCSIR) Complex, Lahore, Pakistan;(2) Department of Chemistry, Govt. College University, Lahore, Pakistan |
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Abstract: | Summary The penicillin G amidase (PGA) activity of a parent strain of E. coli (PCSIR-102) was enhanced by chemical mutagenization with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG). After screening and optimization, a penicillinase deficient mutant (MNNG-37) was isolated and found effective for the production of penicillin G amidase as compared to the parent strain of E. coli (PCSIR-102). Penicillin G amidase activity of MNNG-37 appeared during an early stage of growth, whereas PCSIR-102 did not exhibit PGA activity due to the presence of penicillinase enzyme which inhibits the activity of enzyme PGA. However, MNNG-37 gave a three-fold increase in enzyme activity (231 IU mg−1) as compared to PCSIR-102 (77 IU mg−1) in medium containing 0.15 and 0.1% concentrations of phenylacetic acid, respectively which was added after 6 h of cultivation. The difference in K m values of the enzyme produced by parent strain PCSIR-102 (0.26 mM) and mutant strain MNNG-37 (0.20 mM) is significant (1.3-fold increase in K m value) which may show the superiority of the latter in terms of better enzyme properties. |
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Keywords: | Escherichia coli N-methyl-N′ -nitro-N-nitrosoguanidine Penicillin G amidase Phenylacetic acid |
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