Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1986 Mar 1;234(2):343–347. doi: 10.1042/bj2340343

Structural and kinetic studies on beta-lactamase K1 from Klebsiella aerogenes.

E L Emanuel, J Gagnon, S G Waley
PMCID: PMC1146571  PMID: 3521585

Abstract

beta-Lactamase K1 from Klebsiella aerogenes 1082E hydrolyses both penicillins and cephalosporins comparably and is inhibited by mercurials but not by cloxacillin. These properties distinguish it from those other beta-lactamases that have been allotted to classes on the basis of their amino sequences. beta-Lactamase K1 has been isolated by affinity chromatography; its composition shows resemblances to class A beta-lactamases. Moreover, the N-terminal sequence is similar to those of class A beta-lactamases: there is about 30% identity over the first 32 residues. Furthermore, a putative active-site octapeptide has been isolated and its sequence is similar to the region around the active-site serine residue in class A beta-lactamases. There is one thiol group in beta-lactamase K1; it is not essential for activity. The pH-dependence of kcat. and kcat./Km for the hydrolysis of benzylpenicillin by beta-lactamase K1 were closely similar, suggesting that the rate-determining step is cleavage of the beta-lactam ring.

Full text

PDF
343

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ambler R. P. The structure of beta-lactamases. Philos Trans R Soc Lond B Biol Sci. 1980 May 16;289(1036):321–331. doi: 10.1098/rstb.1980.0049. [DOI] [PubMed] [Google Scholar]
  2. Bibring T., Baxandall J. Peptide analysis by isoelectric focusing in polyacrylamide gels. Anal Biochem. 1978 Mar;85(1):1–14. doi: 10.1016/0003-2697(78)90266-x. [DOI] [PubMed] [Google Scholar]
  3. Bicknell R., Emanuel E. L., Gagnon J., Waley S. G. The production and molecular properties of the zinc beta-lactamase of Pseudomonas maltophilia IID 1275. Biochem J. 1985 Aug 1;229(3):791–797. doi: 10.1042/bj2290791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Bruton C. J., Hartley B. S. Chemical studies on methionyl-tRNA synthetase from Escherichia coli. J Mol Biol. 1970 Sep 14;52(2):165–178. doi: 10.1016/0022-2836(70)90023-9. [DOI] [PubMed] [Google Scholar]
  6. Bush K., Freudenberger J. S., Sykes R. B. Interaction of azthreonam and related monobactams with beta-lactamases from gram-negative bacteria. Antimicrob Agents Chemother. 1982 Sep;22(3):414–420. doi: 10.1128/aac.22.3.414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Campbell D. G., Gagnon J., Reid K. B., Williams A. F. Rat brain Thy-1 glycoprotein. The amino acid sequence, disulphide bonds and an unusual hydrophobic region. Biochem J. 1981 Apr 1;195(1):15–30. doi: 10.1042/bj1950015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cartwright S. J., Waley S. G. Purification of beta-lactamases by affinity chromatography on phenylboronic acid-agarose. Biochem J. 1984 Jul 15;221(2):505–512. doi: 10.1042/bj2210505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Connolly A. K., Waley S. G. Characterization of the membrane beta-lactamase in Bacillus cereus 569/H/9. Biochemistry. 1983 Sep 27;22(20):4647–4651. doi: 10.1021/bi00289a006. [DOI] [PubMed] [Google Scholar]
  10. Cornish-Bowden A. Estimation of the dissociation constants of enzyme-substrate complexes from steady-state measurements. Interpretation of pH-independence of Km. Biochem J. 1976 Feb 1;153(2):455–461. doi: 10.1042/bj1530455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Davies R. B., Abraham E. P. Metal cofactor requirements of beta-lactamase II. Biochem J. 1974 Oct;143(1):129–135. doi: 10.1042/bj1430129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. HAMILTON-MILLER J. M. Penicillinase from Klebsiella aerogenes. A comparison with penicillinases from gram-positive species. Biochem J. 1963 Apr;87:209–214. doi: 10.1042/bj0870209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hardy L. W., Kirsch J. F. pH dependence and solvent deuterium oxide kinetic isotope effects on Bacillus cereus beta-lactamase I catalyzed reactions. Biochemistry. 1984 Mar 13;23(6):1282–1287. doi: 10.1021/bi00301a041. [DOI] [PubMed] [Google Scholar]
  14. Holland S., Dale J. W. Improved purification and characterization of the OXA-2 beta-lactamase. Biochem J. 1984 Dec 15;224(3):1009–1013. doi: 10.1042/bj2241009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jack G. W. The purification and some properties of a -lactamase sensitive to inhibition by p-chloromercuribenzoate. Biochim Biophys Acta. 1971 Nov 13;250(2):428–436. doi: 10.1016/0005-2744(71)90199-9. [DOI] [PubMed] [Google Scholar]
  16. Jaurin B., Grundström T. ampC cephalosporinase of Escherichia coli K-12 has a different evolutionary origin from that of beta-lactamases of the penicillinase type. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4897–4901. doi: 10.1073/pnas.78.8.4897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Joris B., De Meester F., Galleni M., Reckinger G., Coyette J., Frere J. M., Van Beeumen J. The beta-lactamase of Enterobacter cloacae P99. Chemical properties, N-terminal sequence and interaction with 6 beta-halogenopenicillanates. Biochem J. 1985 May 15;228(1):241–248. doi: 10.1042/bj2280241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Joris B., Dusart J., Frere J. M., van Beeumen J., Emanuel E. L., Petursson S., Gagnon J., Waley S. G. The active site of the P99 beta-lactamase from Enterobacter cloacae. Biochem J. 1984 Oct 1;223(1):271–274. doi: 10.1042/bj2230271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Knott-Hunziker V., Petursson S., Waley S. G., Jaurin B., Grundström T. The acyl-enzyme mechanism of beta-lactamase action. The evidence for class C Beta-lactamases. Biochem J. 1982 Nov 1;207(2):315–322. doi: 10.1042/bj2070315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marshall M. J., Ross G. W., Chanter K. V., Harris A. M. Comparison of the substrate specificities of the -lactamases from Klebsiella aerogenes 1082E and Enterobacter cloacae P99. Appl Microbiol. 1972 Apr;23(4):765–769. doi: 10.1128/am.23.4.765-769.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Richmond M. H., Sykes R. B. The beta-lactamases of gram-negative bacteria and their possible physiological role. Adv Microb Physiol. 1973;9:31–88. doi: 10.1016/s0065-2911(08)60376-8. [DOI] [PubMed] [Google Scholar]
  22. Richmond M. H. The beta-lactamase stability of a novel beta-lactam antibiotic containing a 7 alpha-methoxyoxacephem nucleus. J Antimicrob Chemother. 1980 Jul;6(4):445–453. doi: 10.1093/jac/6.4.445. [DOI] [PubMed] [Google Scholar]
  23. Ross G. W., Boulton M. G. Purification of beta-lactamases on QAE-sephadex. Biochim Biophys Acta. 1973 Jun 6;309(2):430–439. doi: 10.1016/0005-2744(73)90041-7. [DOI] [PubMed] [Google Scholar]
  24. Selwyn M. J. A simple test for inactivation of an enzyme during assay. Biochim Biophys Acta. 1965 Jul 29;105(1):193–195. doi: 10.1016/s0926-6593(65)80190-4. [DOI] [PubMed] [Google Scholar]
  25. Sloma A., Gross M. Molecular cloning and nucleotide sequence of the type I beta-lactamase gene from Bacillus cereus. Nucleic Acids Res. 1983 Jul 25;11(14):4997–5004. doi: 10.1093/nar/11.14.4997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sykes R. B. The classification and terminology of enzymes that hydrolyze beta-lactam antibiotics. J Infect Dis. 1982 May;145(5):762–765. doi: 10.1093/infdis/145.2.762. [DOI] [PubMed] [Google Scholar]
  27. Waley S. G. A spectrophotometric assay of beta-lactamase action on penicillins. Biochem J. 1974 Jun;139(3):789–790. doi: 10.1042/bj1390789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Waley S. G. The pH-dependence and group modification of beta-lactamase I. Biochem J. 1975 Sep;149(3):547–551. doi: 10.1042/bj1490547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  30. Wharton C. W., Szawelski R. J. Half-time analysis of the integrated Michaelis equation. Simulation and use of the half-time plot and its direct linear variant in the analysis of some alpha-chymotrypsin, papain- and fumarase-catalysed reactions. Biochem J. 1982 May 1;203(2):351–360. doi: 10.1042/bj2030351. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES