Sharif Digital Repository

Immobilization of enzymes on nonporous supports provides a suitable model for investigating the effect of external mass transfer limitation on the reaction rate in the absence of internal diffusional resistance. In this study, deacylation of penicillin G was investigated using penicillin acylase immobilized on ultrafine silica particles. Kinetic studies were performed within the low-substrate-concentration region, where the external mass transfer limitation becomes significant. To predict the apparent kinetic parameters and the overall effectiveness factor, knowledge of the external mass transfer coefficient, kLa, is necessary. Although various correlations exist for estimation of kLa, in... 

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... 

The kinetics of the enzymic reaction of penicillin G acylase from a mutant of Escherichia coli ATCC 11105 in forward and reverse directions were studied and the kinetic constants determined. Results show that the enzyme is inhibited by excess substrate, penicillin G (Pen G), and by both products. The non-competitive inhibition by 6-aminopenicillanic acid (6-APA) and competitive inhibition by phenylacetic acid were observed for the ordered uni bi deacylation reaction in the forward direction. The optimum pH value for the reverse acylation reaction was 5.7. The bi uni mechanism for the reverse reaction was investigated and the inhibitory effects of the substrates, 6-APA and phenyl acetic acid,...