1887

Abstract

SUMMARY: -Lactamases (EC. 3.5.2.6) from strains of Gram-negative bacteria have been studied using analytical isoelectric focusing. This permits a visual comparison of the patterns of -lactamase bands produced by enzymes from different organisms. Purification of crude intracellular preparations is unnecessary and the technique is sufficiently sensitive to demonstrate -lactamase in mutants previously reported to lack the enzyme. R factor RTEM and RP1 -lactamases that have not been distinguished from one another biochemically or immunologically can be differentiated by isoelectric focusing. Conversely, the enzymes specified by the R factors RTEM, R1 and R, with identical isoelectric focusing patterns, have the same biochemical properties. Chromosomal and R factor-mediated -lactamases from single strains have been separated and their identities confirmed by immunoisoelectric focusing. R factor-mediated enzymes gave identical isoelectric focusing patterns irrespective of the host strain. Isoelectric focusing can therefore be used to observe the transfer of -lactamases carried by R factors.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-88-1-169
1975-05-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/88/1/mic-88-1-169.html?itemId=/content/journal/micro/10.1099/00221287-88-1-169&mimeType=html&fmt=ahah

References

  1. Awdeh Z. L., Williamson A. R., Askonas B. A. 1968; Isoelectric focusing in polyacrylamide gel and its application to immunoglobulins. Nature; London: 21966–67
    [Google Scholar]
  2. Awdeh Z. L., Williamson A. R., Askonas B. A. 1970; One cell-one immunoglobulin: origin of limited heterogeneity of myeloma proteins. Biochemical Journal 116:241–248
    [Google Scholar]
  3. Boman H. G., Eriksson-Grennberg K. G., Normark S., Matsson E. 1968; Resistance of Escherichia coli to penicillins. IV. Genetic study of mutants resistant to d,l-ampicillin concentrations of 100 μg/ml. Genetical Research 12:169–185
    [Google Scholar]
  4. Burman L. G., NordströM K., Boman H. G. 1968; Resistance of Escherichia coli to penicillins. V. Physiological comparison of two isogenic strains, one with chromosomally and one with episomally mediated ampicillin resistance. Journal of Bacteriology 96:438–446
    [Google Scholar]
  5. Burman L. G., Park J. T., Lindström E. B., Boman H. G. 1973; Resistance of Escherichia coli to penicillins: identification of the structural gene for the chromosomal penicillinase. Journal of Bacteriology 116:123–130
    [Google Scholar]
  6. CarlströM A., Vesterberg O. 1967; Isoelectric focusing and separation of the subcomponents of lactoperoxidase. Acta chemica scandinavica 21:271–278
    [Google Scholar]
  7. Cowan S. T., Steel K. J. 1965 Identification of Medical Bacteria pp. 76–82 Cambridge: Cambridge University Press;
    [Google Scholar]
  8. Dale J. W. 1971; Characterization of the β-lactamase specified by the resistance factor R1818 in E. coli K-12 and other Gram-negative bacteria. Biochemical Journal 123:501–505
    [Google Scholar]
  9. Dale J. W., Smith J. T. 1971; Some relationships between R-factor and chromosomal β-lactamases in Gram-negative bacteria. Biochemical Journal 123:507–512
    [Google Scholar]
  10. Dale J. W., Smith J. T. 1972; A direct comparison of two unusual R-factor-mediated β-lactamases. Biochemical Journal 128:173–174
    [Google Scholar]
  11. Datta N. 1962; Transmissible drug resistance in an epidemic strain of Salmonella typhimurium. Journal of Hygiene 60:301–310
    [Google Scholar]
  12. Datta N., Kontomichalou P. 1965; Penicillinase synthesis controlled by infectious R-factors in Enterobacteriaceae. Nature; London: 208239–241
    [Google Scholar]
  13. Fleming P. C., Goldner M., Glass D. G. 1963; Observations on the nature, distribution and significance of cephalosporinase. Lancet i:1399–1401
    [Google Scholar]
  14. Frater R. 1970; Artifacts in isoelectric focusing. Journal of Chromatography 50:469–474
    [Google Scholar]
  15. Frère J.-M., Ghuysen J.-M., Reynolds P. E., Moreno R., Perkins H. R. 1974; Binding of β-lactam antibiotics to the exocellular dd-carboxypeptidase-transpeptidase of Streptomyces R39. Biochemical Journal 173:241–249
    [Google Scholar]
  16. Goldner M., Glass D. G., Fleming P. C. 1969; Spontaneous mutant with loss of β-lactamase in Aerobacter cloacae. Journal of Bacteriology 97:961
    [Google Scholar]
  17. Haglund H. 1967; Isoelectric focusing in natural pH gradients - a technique of growing importance for fractionation and characterization of proteins. Science Tools 14:17–23
    [Google Scholar]
  18. Hamilton-Miller J. M. T. 1967; Hydrolysis of cephalosporins by β-lactamases from Gram-negative bacteria. Nature; London: 2141333–1334
    [Google Scholar]
  19. Hennessey T. D. G. 1967; Inducible β-lactamase in Enterobacter. Journal of General Microbiology 49:277–285
    [Google Scholar]
  20. Illingworth J. A. 1972; Anomalous behaviour of yeast isocitrate dehydrogenase during isoelectric focusing. Biochemical Journal 129:1125–1130
    [Google Scholar]
  21. Jack G. W., Richmond M. H. 1970; A comparative study of eight distinct β-lactamases synthesized by Gram-negative bacteria. Journal of General Microbiology 61:43–61
    [Google Scholar]
  22. Marshall M. J., Ross G. W., Chanter K. V., Harris A. M. 1972; Comparison of the substrate specificities of the β-lactamases from Klebsiella aerogenes 1082E and Enterobacter cloacae P99. Applied Microbiology 23:765–769
    [Google Scholar]
  23. Meynell E., Datta N. 1966; The relation of resistance transfer factors to the R-factor (sex-factor) of Escherichia coli k12. Genetical Research 7:134–140
    [Google Scholar]
  24. Newsom S. W. B., Marshall M. J., Harris A. M. 1974; Enterobacteria, β-lactam antibiotics and β-lactamases in clinical practice. Journal of Medical Microbiology 7:473–482
    [Google Scholar]
  25. Nguyen-Distèche M., Pollock J. J., Ghuysen J.-M., Puig J., Reynolds P., Perkins H. R., Coyette J., Salton M. R. J. 1973; Sensitivity to ampicillin and cephalothin of enzymes involved in wall peptide cross-linking in Escherichia coli KI2, strain 44. European Journal of Biochemistry 41:457–463
    [Google Scholar]
  26. O’Callaghan C. H., Morris A. 1972; Inhibition of β-lactamases by β-lactam antibiotics. Antimicrobial Agents and Chemotherapy 2:442–448
    [Google Scholar]
  27. O’Callaghan C. H., Morris A., Kirby S. M., Shingler A. H. 1972; Novel method for detection of β-lactamases by using a chromogenic cephalosporin substrate. Antimicrobial Agents and Chemotherapy 1:283–288
    [Google Scholar]
  28. Perret C. J. 1954; Iodometric assay of penicillinase. Nature; London: 1741012–1013
    [Google Scholar]
  29. Richmond M. H., Sykes R. B. 1972; The chromosomal integration of a β-lactamase gene derived from the P-type R-factor RP1 in Escherichia coli. Genetical Research 20:231–237
    [Google Scholar]
  30. Richmond M. H., Sykes R. B. 1973; The β-lactamases of Gram-negative bacteria and their possible nhvsioloeical role. Advances in Microbial Phvsiology 9:31–88
    [Google Scholar]
  31. Ross G. W., Boulton M. G. 1972; Improvement of the specificity of an antiserum to β-lactamase by absorption with a mutant which does not produce the enzyme. Journal of Bacteriology 112:1435–1437
    [Google Scholar]
  32. Ross G. W., Boulton M. G. 1973; Purification of β-lactamases on QAE-Sephadex. Biochimica et biophysica acta 309:430–439
    [Google Scholar]
  33. Salaman M. R., Williamson A. R. 1971; Isoelectric focusing of proteins in the native and denatured states. Biochemical Journal 122:93–99
    [Google Scholar]
  34. Sawai T., Mitsuhashi S., Yamagishi S. 1968; Drug resistance of enteric bacteria. XIV. Comparison of β-lactamases in Gram-negative rod bacteria resistant to α-aminobenzyl penicillin. Japanese Journal of Microbiology 12:423–434
    [Google Scholar]
  35. Smith J. T. 1969; R-factor gene expression in Gram-negative bacteria. Journal of General Microbiology 55:109–120
    [Google Scholar]
  36. Sykes R. B., Richmond M. H. 1970; Intergeneric transfer of a β-lactamase gene between Ps. aeruginosa and E. coli. Nature; London: 226:952–954
    [Google Scholar]
  37. Yamagishi S., O’hara K., Sawai T., Mitsuhashi S. 1969; The purification and properties of penicillin β-lactamases mediated by transmissible R-factors in Escherichia coli. Journal of Biochemistry 66:11–20
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-88-1-169
Loading
/content/journal/micro/10.1099/00221287-88-1-169
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error