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PMID: 3933490 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Single-turnover and steady-state kinetics of hydrolysis of cephalosporins by beta-lactamase I from Bacillus cereus.

The Biochemical journal ·Vol. 231 ·No. 1 ·1985-10-01 ·Pages 83-8

Bicknell R, Waley SG

Abstract

The kinetics of the hydrolysis of two cephalosporins by beta-lactamase I from Bacillus cereus 569/H/9 has been studied by single-turnover and steady-state methods. Single-turnover kinetics could be measured over the time scale of minutes when cephalosporin C was the substrate. The other substrate, 7-(2',4'-dinitrophenylamino)deacetoxycephalosporanic acid, was hydrolysed even more slowly, and has potential for use in crystallographic studies of beta-lactamases. Comparison of single-turnover and steady-state kinetics showed that, for both substrates, opening the beta-lactam ring (i.e. acylation of the enzyme) was the rate-determining step. Thus the non-covalent enzyme-substrate complex is expected to be the intermediate observed crystallographically.

MeSH Terms
Ammonium Sulfate/pharmacology Bacillus cereus/enzymology Cephalosporins/metabolism Hydrolysis Kinetics Penicillinase/metabolism Spectrophotometry
Chemicals
Cephalosporins Penicillinase Ammonium Sulfate
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Bicknell R
Waley S G
References (36)
36 references, click to expand
  1. Purification and properties of thiol beta-lactamase. A mutant of pBR322 beta-lactamase in which the active site serine has been replaced with cysteine.
    J Biol Chem. 1984 Apr 25;259(8):5327-32 PMID: 6425288
  2. Crystallographic data for the beta-lactamase from Enterobacter cloacae P99.
    J Mol Biol. 1983 Dec 5;171(2):237-8 PMID: 6606712
  3. Active site of staphylococcal beta-lactamase.
    Philos Trans R Soc Lond B Biol Sci. 1980 May 16;289(1036):370-2 PMID: 6109335
  4. Mechanism of substrate-induced inactivation of beta-lactamase I.
    Eur J Biochem. 1980 Aug;109(2):575-80 PMID: 6773776
  5. Separation, purification and properties of beta-lactamase I and beta-lactamase II from Bacillus cereus 569/H/9.
    Biochem J. 1974 Oct;143(1):115-27 PMID: 4219278
  6. An easy method for the determination of initial rates.
    Biochem J. 1981 Mar 1;193(3):1009-12 PMID: 7305952
  7. 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):4897-901 PMID: 6795623
  8. Production of a variant of beta-lactamase II with selectively decreased cephalosporinase activity by a mutant of Bacillus cereus 569/H/9.
    Biochem J. 1980 Oct 1;191(1):111-6 PMID: 6781486
  9. beta-Lactamase proceeds via an acyl-enzyme intermediate. Interaction of the Escherichia coli RTEM enzyme with cefoxitin.
    Biochemistry. 1980 Jun 24;19(13):2895-901 PMID: 6994800
  10. Isolation of a covalent intermediate in beta -lactamase I catalysis.
    FEBS Lett. 1982 Jan 25;137(2):186-8 PMID: 6800840
  11. Oligonucleotide-directed mutagenesis as a general and powerful method for studies of protein function.
    Proc Natl Acad Sci U S A. 1982 Nov;79(21):6409-13 PMID: 6983070
  12. A simple test for inactivation of an enzyme during assay.
    Biochim Biophys Acta. 1965 Jul 29;105(1):193-5 PMID: 4221326
  13. Interaction between serine beta-lactamases and class A substrates: a kinetic analysis and a reaction pathway hypothesis.
    Biochem Pharmacol. 1981 Mar 15;30(6):549-52 PMID: 6973976
  14. Proceedings of the biochemical society.
    Biochem J. 1966 Jan;98(1):1-16P PMID: 16749173
  15. Penicillinase active sites: labelling of serine-44 in beta-lactamase I by 6beta-bromopenicillanic acid.
    FEBS Lett. 1979 Mar 1;99(1):59-61 PMID: 220094
  16. Inactivation of Bacillus cereus beta-lactamase I by 6 beta-bromopencillanic acid: mechanism.
    Biochemistry. 1980 Aug 19;19(17):3996-4003 PMID: 6773559
  17. Inactivation of the RTEM beta-lactamase from Escherichia coli. Interaction of penam sulfones with enzyme.
    Biochemistry. 1981 May 12;20(10):2726-31 PMID: 7018564
  18. The active site of the P99 beta-lactamase from Enterobacter cloacae.
    Biochem J. 1984 Oct 1;223(1):271-4 PMID: 6333871
  19. The acyl-enzyme mechanism of beta-lactamase action. The evidence for class C Beta-lactamases.
    Biochem J. 1982 Nov 1;207(2):315-22 PMID: 6818947
  20. Purification and properties of inducible penicillin beta-lactamase isolated from Pseudomonas maltophilia.
    Antimicrob Agents Chemother. 1982 Oct;22(4):564-70 PMID: 6983856
  21. Acquisition of substrate-specific parameters during the catalytic reaction of penicillinase.
    Proc Natl Acad Sci U S A. 1976 Apr;73(4):1048-52 PMID: 817286
  22. 5-5A crystallographic structure of penicillin beta-lactamase and radius of gyration in solution.
    J Mol Biol. 1976 Jul 15;104(4):865-75 PMID: 785016
  23. Estimation of Michaelis constant and maximum velocity from the direct linear plot.
    Biochim Biophys Acta. 1978 Mar 14;523(1):268-72 PMID: 629990
  24. Isolation and properties of an inducible and a constitutive beta-lactamase from Pseudomonas aeruginosa.
    J Gen Microbiol. 1982 Jan;128(1):155-9 PMID: 6806437
  25. Behaviour of some derivatives of 7-aminocephalosporanic acid and 6-aminopenicillanic acidas substrates, inhibitors and inducers of penicillinases.
    Biochem J. 1962 Apr;83:52-63 PMID: 13882319
  26. Preliminary crystallographic data for beta-lactamase I from Bacillus cereus 569.
    J Mol Biol. 1978 Apr 15;120(3):447-9 PMID: 418181
  27. Pre-steady state beta-lactamase kinetics. Observation of a covalent intermediate during turnover of a fluorescent cephalosporin by the beta-lactamase of STaphylococcus aureus PC1.
    J Biol Chem. 1981 Nov 25;256(22):11401-4 PMID: 6975275
  28. The structure of beta-lactamases.
    Philos Trans R Soc Lond B Biol Sci. 1980 May 16;289(1036):321-31 PMID: 6109327
  29. The crystal structure of beta-lactamase from Staphylococcus aureus at 0.5 nm resolution.
    Biochem J. 1985 Jan 1;225(1):167-76 PMID: 2983660
  30. Thiol-beta-lactamase: replacement of the active-site serine of RTEM beta-lactamase by a cysteine residue.
    Proc Natl Acad Sci U S A. 1982 Dec;79(23):7157-60 PMID: 6818541
  31. 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):351-60 PMID: 7115291
  32. Beta-lactamase inactivation by mechanism-based reagents.
    Philos Trans R Soc Lond B Biol Sci. 1980 May 16;289(1036):309-19 PMID: 6109326
  33. The ficin-catalysed hydrolysis of p-nitrophenyl hippurate. Detailed kinetics including the measurement of the apparent dissociation constant for the enzyme-substrate complex.
    FEBS Lett. 1969 Aug;4(4):299-306 PMID: 11947209
  34. Crystallization and preliminary X-ray data for the exocellular beta-lactamase of Bacillus licheniformis 749/C.
    J Mol Biol. 1985 Jan 5;181(1):145-6 PMID: 3872374
  35. A comparison of the action of penicillinase on benzylpenicillin and cephalosporin N and the competitive inhibition of penicillinase by cephalosporin C.
    Biochem J. 1956 Aug;63(4):628-34 PMID: 13355861
  36. Pre-steady state beta-lactamase kinetics. The trapping of a covalent intermediate and the interpretation of pH rate profiles.
    J Biol Chem. 1983 Nov 10;258(21):13120-6 PMID: 6605346
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2310083
Published
1985-10-01
Pages
83-8
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1152706
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2310083 PMC Full Text View journal details
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