Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1973 Sep;115(3):1045–1054. doi: 10.1128/jb.115.3.1045-1054.1973

Penicillinases of Klebsiella pneumoniae and Their Phylogenetic Relationship to Penicillinases Mediated by R Factors

Tetsuo Sawai 1, Saburo Yamagishi 1, Susumu Mitsuhashi 1
PMCID: PMC246352  PMID: 4199503

Abstract

On the assumption that the penicillinase determinants on a group of R factors conferring ampicillin resistance have a phylogenetically close relationship to the penicillinase gene of the Klebsiella group, the penicillinases from four strains of K. pneumoniae, GN69, GN1103R−, GN422, and GN118, were purified 230- to 1,000-fold and compared with the known two R-factor-mediated penicillinases. By gel filtration on Sephadex G-75, the molecular weights were estimated to be 17,400, 18,100, 20,000 and 18,300, respectively, which are slightly lower than those of the R-factor penicillinases. The isoelectric points of the Klebsiella penicillinases were not in agreement with those of the R-factor penicillinases. All the enzymes showed a pH optimum between 6.3 to 7.2 and a temperature optimum of 45 C, and those properties, together with behavior towards inhibitors, were about the same as those in the R-factor penicillinases. The substrate specificity and the Michaelis constants of the Klebsiella penicillinases for penicillins and cephaloridine were broadly similar to those of the R-factor penicillinases, however, some variations were found even among the four penicillinases of K. pneumoniae. The reactivities of the four penicillinases of K. pneumoniae with the antiserum against one R-factor penicillinase were tested, and three of the four Klebsiella penicillinases were found to be indistinguishable immunologically from both R-factor penicillinases. The remaining Klebsiella penicillinase, from GN1103R−, showed an immunological partial homology with the R-factor penicillinases.

Full text

PDF
1052

Images in this article

1050Image
on p.1050

Selected References

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

  1. ANDERSON E. S., DATTA N. RESISTANCE TO PENICILLINS AND ITS TRANSFER IN ENTEROBACTERIACEAE. Lancet. 1965 Feb 20;1(7382):407–409. doi: 10.1016/s0140-6736(65)90004-8. [DOI] [PubMed] [Google Scholar]
  2. AYLIFFE G. A. INDUCTION OF CEPHALOSPORINASE AND PENICILLINASE IN PROTEUS SPECIES. Nature. 1964 Mar 7;201:1032–1032. doi: 10.1038/2011032a0. [DOI] [PubMed] [Google Scholar]
  3. Anderson E. S. Origin of transferable drug-resistance factors in the enterobacteriaceae. Br Med J. 1965 Nov 27;2(5473):1289–1291. doi: 10.1136/bmj.2.5473.1289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anderson E. S. The ecology of transferable drug resistance in the enterobacteria. Annu Rev Microbiol. 1968;22:131–180. doi: 10.1146/annurev.mi.22.100168.001023. [DOI] [PubMed] [Google Scholar]
  5. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ayliffe G. A. Cephalosporinase and penicillinase activity of Gram-negative bacteria. J Gen Microbiol. 1965 Jul;40(1):119–126. doi: 10.1099/00221287-40-1-119. [DOI] [PubMed] [Google Scholar]
  7. Dale J. W., Smith J. T. Some relationships between R-factor and chromosomal -lactamase in Gram-negative bacteria. Biochem J. 1971 Jul;123(4):507–512. doi: 10.1042/bj1230507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Datta N., Richmond M. H. The purification and properties of a penicillinase whose synthesis is mediated by an R-factor in Escherichia coli. Biochem J. 1966 Jan;98(1):204–209. doi: 10.1042/bj0980204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. FLEMING P. C., GOLDNER M., GLASS D. G. Observations on the nature, distribution, and significance of cephalosporinase. Lancet. 1963 Jun 29;1(7296):1399–1401. doi: 10.1016/s0140-6736(63)92051-8. [DOI] [PubMed] [Google Scholar]
  10. Fullbrook P. D., Elson S. W., Slocombe B. R-factor mediated beta-lactamase in Pseudomonas aeruginosa. Nature. 1970 Jun 13;226(5250):1054–1056. doi: 10.1038/2261054a0. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Hamilton-Miller J. M., Smith J. T., Knox R. Interaction of cephaloridine with penicillinase-producing gram-negative bacteria. Nature. 1965 Oct 16;208(5007):235–237. doi: 10.1038/208235a0. [DOI] [PubMed] [Google Scholar]
  13. Hamilton-miller J. M.T. Wild type variants of penicillinase from Klebsiella aerogenes. FEBS Lett. 1968 Aug;1(2):86–88. doi: 10.1016/0014-5793(68)80026-2. [DOI] [PubMed] [Google Scholar]
  14. Hennessey T. D. Inducible beta-lactamase in Enterobacter. J Gen Microbiol. 1967 Nov;49(2):277–285. doi: 10.1099/00221287-49-2-277. [DOI] [PubMed] [Google Scholar]
  15. Jack G. W., Richmond M. H. A comparative study of eight distinct beta-lactamases synthesized by gram-negative bacteria. J Gen Microbiol. 1970 Apr;61(1):43–61. doi: 10.1099/00221287-61-1-43. [DOI] [PubMed] [Google Scholar]
  16. Kameda M., Harada K., Suzuki M., Mitshuhasi S. Formation of transferable drug resistance factor by recombination between resistance determinants and transfer factors. Jpn J Microbiol. 1969 Sep;13(3):255–262. doi: 10.1111/j.1348-0421.1969.tb00465.x. [DOI] [PubMed] [Google Scholar]
  17. Kontomichalou P., Mitani M., Clowes R. C. Circular R-factor molecules controlling penicillinase synthesis, replicating in Escherichia coli under either relaxed or stringent control. J Bacteriol. 1970 Oct;104(1):34–44. doi: 10.1128/jb.104.1.34-44.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  19. Lindqvist R. C., Nordström K. Resistance of Escherichia coli to penicillins. VII. Purification and characterization of a penicillinase mediated by the R factor R1. J Bacteriol. 1970 Jan;101(1):232–239. doi: 10.1128/jb.101.1.232-239.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Linström E. B., Boman H. G., Steele B. B. Resistance of Escherichia coli to penicillins. VI. Purification and characterization of the chromosomally mediated penicillinase present in ampA-containing strains. J Bacteriol. 1970 Jan;101(1):218–231. doi: 10.1128/jb.101.1.218-231.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Matsumoto H., Sawai T., Tazaki T., Yamagishi S., Mitsuhashi S. Characterization of the chromosomally mediated penicillinase in Klebsiella pneumoniae. Jpn J Microbiol. 1972 May;16(3):169–176. doi: 10.1111/j.1348-0421.1972.tb00645.x. [DOI] [PubMed] [Google Scholar]
  22. Matsumoto H., Tazaki T. Genetic mapping of aro, pyr, and pur markers in Klebsiella pneumoniae. Jpn J Microbiol. 1971 Jan;15(1):11–20. [PubMed] [Google Scholar]
  23. Matsumoto H., Tazaki T. Genetic recombination in Klebsiella pneumoniae. An approach to genetic linkage mapping. Jpn J Microbiol. 1970 Mar;14(2):129–141. doi: 10.1111/j.1348-0421.1970.tb00500.x. [DOI] [PubMed] [Google Scholar]
  24. Meynell E., Meynell G. G., Datta N. Phylogenetic relationships of drug-resistance factors and other transmissible bacterial plasmids. Bacteriol Rev. 1968 Mar;32(1):55–83. doi: 10.1128/br.32.1.55-83.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mitsuhashi S., Kameda M., Harada K., Suzuki M. Formation of recombinants between nontransmissible drug-resistance determinants and transfer factors. J Bacteriol. 1969 Mar;97(3):1520–1521. doi: 10.1128/jb.97.3.1520-1521.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. NOVICK R. P. Micro-iodometric assay for penicillinase. Biochem J. 1962 May;83:236–240. doi: 10.1042/bj0830236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nisioka T., Mitani M., Clowes R. Composite circular forms of R factor deoxyribonucleic acid molecules. J Bacteriol. 1969 Jan;97(1):376–385. doi: 10.1128/jb.97.1.376-385.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. PERRET C. J. Iodometric assay of penicillinase. Nature. 1954 Nov 27;174(4439):1012–1013. doi: 10.1038/1741012a0. [DOI] [PubMed] [Google Scholar]
  29. SMITH J. T., HAMILTON-MILLER J. M. Differences between pencillinases from gram-positive and gram-negative bacteria. Nature. 1963 Mar 9;197:976–978. doi: 10.1038/197976a0. [DOI] [PubMed] [Google Scholar]
  30. SMITH J. T. Penicillinase and ampicillin resistance in a strain of Escherichia coli. J Gen Microbiol. 1963 Feb;30:299–306. doi: 10.1099/00221287-30-2-299. [DOI] [PubMed] [Google Scholar]
  31. Sabath L. D., Abraham E. P. Zinc as a cofactor for cephalosporinase from Bacillus cereus 569. Biochem J. 1966 Jan;98(1):11C–13C. doi: 10.1042/bj0980011c. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sawai T., Mitsuhashi S., Yamagishi S. Drug resistance of enteric bacteria. XIV. Comparison of beta-lactamases in gram-negative rod bacteria resistant to alpha-aminobenzylpenicillin. Jpn J Microbiol. 1968 Dec;12(4):423–434. doi: 10.1111/j.1348-0421.1968.tb00415.x. [DOI] [PubMed] [Google Scholar]
  33. Sawai T., Takahashi K., Yamagishi S., Mitsuhashi S. Variant of penicillinase mediated by an R factor in Escherichia coli. J Bacteriol. 1970 Nov;104(2):620–629. doi: 10.1128/jb.104.2.620-629.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Silver R. P., Falkow S. Studies on resistance transfer factor deoxyribonucleic acid in Escherichia coli. J Bacteriol. 1970 Oct;104(1):340–344. doi: 10.1128/jb.104.1.340-344.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  36. Yamagishi S., O'Hara K., Sawai T., Mitsuhashi S. The purification and properties of penicillin beta-lactamases mediated by transmissible R factors in Escherichia coli. J Biochem. 1969 Jul;66(1):11–20. doi: 10.1093/oxfordjournals.jbchem.a129111. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES