Purification and characterization of an extracellular β-lactamase produced by Free

Abstract

A β-lactamase was purified 430-fold from the culture supernatant of by ion exchange chromatography on CM-Sephadex and affinity chromatography on phenylboronic-acid-agarose. The purified enzyme was homogeneous as judged by SDS-PAGE, and was characterized with respect to molecular mass (38 and 41 kDa by gel filtration on Sephadex G-75 and SDS-PAGE, respectively), pH optimum (pH 7·0), temperature optimum (45 °C) and isoelectric point (9·3). The β-lactamase showed mainly cephalosporinase activity. It was inhibited by cloxacillin, carbenicillin, penicillanic acid sulphone (sulbactam) and aztreonam. It was not inhibited by clavulanic acid up to a concentration of 0·25 mM. Neither EDTA nor -chlormercuribenzoate, up to concentrations of 1 or 100 mM, respectively, affected activity. According to these characteristics, it is a typical CEP-N cephalosporinase.

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1992-06-01
2024-03-28
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References

  1. Bauernfeind A. 1986; Classification of beta-lactamases. Reviews of Infectious Diseases 8:Suppl.470–481
    [Google Scholar]
  2. Bergogne-Bérézin E., Joly-Guillou M. L., Vieu J. F. 1987; Epidemiology of nosocomial infections due to Acinetobaeter calco-acetkus . Journal of Hospital Infection 10:105–113
    [Google Scholar]
  3. Blechschmidt B., Borneleit P., Emanouilidis I., Lehmann W., Kleber H.-P. 1989; Extracellular location of a /β-lactamase produced by Acinetobaeter calcoaceticus . Applied Microbiology and Biotechnology 32:85–89
    [Google Scholar]
  4. Blechschmidt B., Borneleit P., Kleber H.-P. 1991; Influence of cultivation and induction conditions on β-lactamase production in Acinetobaeter calcoaceticus . Applied Microbiologv and Biotechnology 36:364–368
    [Google Scholar]
  5. Borneleit P., Blechschmidt B., Eschrich K., Kleber H.-P. 1991; β-Lactamase export across the outer membrane of Acinetobaeter calcoaceticus : perturbation of the outer membrane lipopoly-saccharide leaflet by enzyme overproduction. Archives of Microbiology 155:500–504
    [Google Scholar]
  6. Bush Κ. 1989; Characterization of β-lactamases. Antimicrobial Agents and Chemotherapy 33:259–263
    [Google Scholar]
  7. Bush K. 1989; Classification of β-lactamases: Groups 1, 2a, 2b, and 2b’. Antimicrobial Agents and Chemotherapy 33:264–270
    [Google Scholar]
  8. Bush K. 1989c; Classification of /β-lactamases : Groups 2c, 2d, 2e, 3, and 4. Antimicrobial Agents and Chemotherapy 33:271–276
    [Google Scholar]
  9. Cartwright S. J., Waley S. G. 1984; Purifications of β-lactamases by affinity chromatography on phenylboronic acid-agarose. Biochemical Journal 221:505–512
    [Google Scholar]
  10. Dijkshoorn L., Michel M. F., Degener J. E. 1987a; Cell envelope protein profiles of Acinetobaeter calcoaceticus strains isolated in hospitals. Journal of Medical Microbiology 23:313–319
    [Google Scholar]
  11. Dijkshoorn L., Van Vianen W., Degener J. E., Michel M. F. 19876; Typing of Acinetobaeter calcoaceticus strains isolated from hospital patients by cell envelope protein profiles. Epidemiology and Infection 99:659–667
    [Google Scholar]
  12. Dijkshoorn L., Wubbels J. L., Beunders A. J., Degener J. E., Boks A. L., Michel M. F. 1989; Use of protein profiles to identify Acinetobaeter calcoaceticus in a respiratory care unit. Journal of Clinical Pathology 42:853–857
    [Google Scholar]
  13. Dombach G. 1989; Isolierung und Charakterisierung einer Hydan-toinase aus Arthrobacter sp. DSM 3747. Dissertation, TU Braunschweig
    [Google Scholar]
  14. Hikida M., Yoshida M., Mitsuhashi S., Inoue M. 1989; Purification and properties of a cephalosporinase from Acinetobaeter calcoaceticus . Journal of Antibiotics 42:123–126
    [Google Scholar]
  15. Hood J., Amyes S. G. B. 1989; A novel method for the identification and distinction of the β-lactamases of the genus Acinetobaeter . Journal of Applied Bacteriology 67:157–163
    [Google Scholar]
  16. Hood J., Amyes S. G. B. 1991; The chromosomal β-lactamases of the genus Acinetobaeter: enzymes which challenge our imagination. The Biology of Acinetobaeter117–132 Towner K. J., Bergogne-Bérézin E., Fewson C. A. New York: Plenum Press;
    [Google Scholar]
  17. Joly-Guillou M. L., Bergogne-Bérézin E. 1986; Distribution of beta-lactamases in clinical strains ot Acinetobaeter calcoaceticus . 26th Interscience Conference on Antimicrobial Agents and Chemotherapy, New Orleans, Louisiana Abstract 202
    [Google Scholar]
  18. Joly-Guillou M. L., Bergogne-Bérézin E., Moreau N. 1987; Enzymatic resistance to beta-lactams and amino-glycosides in A cinetobacter calcoaceticus . Journal of A ntimicrobial Chemotherapy 20:771–782
    [Google Scholar]
  19. Joly-Guillou M. L., Vallée E., Bergogne-Bérézin E., Phillipon A. 1988; Distribution of beta-lactamases and pheno-type analysis in clinical strains of Acinetobaeter calcoaceticus . Journal of Antimicrobial Chemotherapy 22:597–604
    [Google Scholar]
  20. Kleber H.-P., Schöpp W., Aurich H. 1973; Verwertung von n-Alkanen durch einen Stamm von Acinetobaeter calcoaceticus . Zeitschrift für Allgemeine Mikrobiologie 13:455–477
    [Google Scholar]
  21. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  22. Medeiros A. A. 1984; β-Lactamases. British Medical Bulletin 40:18–27
    [Google Scholar]
  23. Medeiros Α. Α., Hare R., Papa E., Adam C., Miller G. H. 1985; Gram-negative bacilli resistant to third generation cephalosporins: β-lactamase characterisation and susceptibility to SCH 34343. Journal of Antimicrobial Chemotherapy 15:119–124
    [Google Scholar]
  24. Morohoshi T., Saito T. 1977; Beta-lactamase and beta-lactam antibiotic resistance in Acinetobaeter anitratus (syn. A. calcoaceticus). Journal of Antibiotics 30:969–973
    [Google Scholar]
  25. Ohya S., Fujii-Kuriyama Y., Yamamoto M., Sugawara S. 1980; Purification and some properties of β-lactamases from Proteus rettgeri and Proteus insconstans . Microbiologv and Immunology 24:815–824
    [Google Scholar]
  26. Perrett C. J. 1954; Iodometric assay of β-lactamase. Nature, London 174:1012
    [Google Scholar]
  27. Richmond M. H., Sykes R. B. 1973; The jS-lactamases of Gram-negative bacteria and their possible physiological role. Advances in Microbial Physiology 9:31–88
    [Google Scholar]
  28. Samuni A. 1975; A direct spectrophotometric assay and determinations of Michaelis constants for the β-lactamase reaction. Analytical Biochemistry 63:17–26
    [Google Scholar]
  29. Sanders C. C., Sanders W. E. Jr 1986; Type I β-lactamases of Gram-negative bacteria: Interaction with β-lactam antibiotics. Journal of Infectious Diseases 154:792–800
    [Google Scholar]
  30. Sawai T., Kanno M., Tsukamoto K. 1982; Characterization of eight beta-lactamases of Gram-negative bacteria. Journal of Bacteriology 152:567–571
    [Google Scholar]
  31. Waley S. G. 1974; A spectrophotometric assay of β-lactamase action on penicillins. Biochemical Journal 139:159–163
    [Google Scholar]
  32. Yang Y., Wu P., Livermore D. M. 1990; Biochemical characterization of a β-lactamase that hydrolyzes penems and carbapenems from two Serratia marcescens isolates. Antimicrobial Agents and Chemotherapy 34:755–758
    [Google Scholar]
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