PMID: 996Sep 1, 1975

The pH-dependence and group modification of beta-lactamase I

The Biochemical Journal
S G Waley

Abstract

The pH-dependence of the kinetic parameters for the hydrolysis of the beta-lactam ring by beta-lactamase I (penicillinase, EC 3.5.2.6) was studied. Benzylpenicillin and ampicillin (6-[D(-)-alpha-aminophenylacetamido]penicillanic acid) were used. Both kcat. and kcat./Km for both substrates gave bell-shaped plots of parameter versus pH. The pH-dependence of kcat./Km for the two substrates gave the same value (8.6) for the higher apparent pK, and so this value may characterize a group on the free enzyme; the lower apparent pK values were about 5(4.85 for benzylpenicillin, 5.4 for ampicillin). For benzylpenicillin both kcat. and kcat./Km depended on pH in exactly the same way. The value of Km for benzylpenicillin was thus independent of pH, suggesting that ionization of the enzyme's catalytically important groups does not affect binding of this substrate. The pH-dependence of kcat. for ampicillin differed, however, presumably because of the polar group in the side chain. The hypothesis that the pK5 group is a carboxyl group was tested. Three reagents that normally react preferentially with carboxyl groups inactivated the enzyme: the reagents were Woodward's reagent K, a water-soluble carbodi-imide, and triethyloxonium fluoroborate....Continue Reading

Citations

Oct 1, 1983·Medicinal Research Reviews·S J Cartwright, S G Waley
Sep 25, 2014·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Soumya MukherjeeSomdatta Ghosh Dey
Feb 2, 1987·European Journal of Biochemistry·L VarettoC Houssier
Jan 1, 1985·Biotechnology & Genetic Engineering Reviews·A Coulson

Related Concepts

Ukapen
Hydrogen-Ion Concentration
Hydrolysis
Van-Pen-G
Exopenicillinase

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