Skip to main content
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1976 May;9(5):727–735. doi: 10.1128/aac.9.5.727

Formation of Beta-Lactamase in Bacteroides fragilis: Cell-Bound and Extracellular Activity

Barbro Olsson 1, Carl-Erik Nord 1, Torkel Wadström 1
PMCID: PMC429612  PMID: 7997

Abstract

Nine strains of Bacteroides fragilis were cultivated in stirred fermentors and tested for their ability to produce β-lactamase. There was a correlation between formation of β-lactamase and high values of the minimal inhibitory concentration against β-lactam antibiotics. B. fragilis strain B70 was used for optimizing the production of β-lactamase. The highest bacterial yield was obtained in a proteose peptone-yeast extract medium. Optimal conditions for growth and β-lactamase production were obtained at 37 C and pH 7.0. The β-lactamase was released into the surrounding medium during the growth period to about 50%. Osmotic shock released about 20% of the total activity, and remaining activity was found in the cytoplasmic fraction. Substrate profile studies on four β-lactamase-producing strains showed that the enzymes were mainly cephalosporinases. They are inhibited by cloxacillin, p-chloromercuribenzoate, and iodine. Analytical isoelectric focusing in polyacrylamide gel gave an isoelectric point of 4.9 ± 0.2 for three of the strains and 5.6 ± 0.2 for one. Comparison with β-lactamases from aerobic gram-negative species with regard to isoelectric points showed no similarities. Also the molecular weight of the β-lactamase from strain B70 of 43,000 indicates that this is a new class of β-lactamase.

Full text

PDF
727

Selected References

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

  1. Anderson J. D., Sykes R. B. Characterisation of a -lactamase obtained from a strain of Bacteroides fragilis resistant to -lactam antibiotics. J Med Microbiol. 1973 May;6(2):201–206. doi: 10.1099/00222615-6-2-201. [DOI] [PubMed] [Google Scholar]
  2. Cerny G., Teuber M. Differential release of periplasmic versus cytoplasmic enzymes from Escherichia coli B by polymixin B. Arch Mikrobiol. 1971;78(2):166–179. doi: 10.1007/BF00424873. [DOI] [PubMed] [Google Scholar]
  3. Dalland E., Hofstad T. Growth of Bacteroides fragilis in continuous culture and in batch cultures at controlled pH. Appl Microbiol. 1974 Nov;28(5):856–860. doi: 10.1128/am.28.5.856-860.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Del Bene V. E., Farrar W. E., Jr Cephalosporinase activity in Bacteroides fragilis. Antimicrob Agents Chemother. 1973 Mar;3(3):369–372. doi: 10.1128/aac.3.3.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dornbusch K., Nord C. E., Olsson B. Antibiotic susceptibility testing of anaerobic bacteria by the standardized disc diffusion method with special reference to bacteroides fragilis. Scand J Infect Dis. 1975;7(1):59–66. doi: 10.3109/inf.1975.7.issue-1.11. [DOI] [PubMed] [Google Scholar]
  6. Evans D. J., Evans D. G., Gorbach S. L. Polymyxin B-Induced Release of Low-Molecular-Weight, Heat-Labile Enterotoxin from Escherichia coli. Infect Immun. 1974 Nov;10(5):1010–1017. doi: 10.1128/iai.10.5.1010-1017.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. GARROD L. P. Sensitivity of four species of bacteroides to antibiotics. Br Med J. 1955 Dec 24;2(4955):1529–1531. doi: 10.1136/bmj.2.4955.1529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gorbach S. L., Bartlett J. G. Anaerobic infections. 1. N Engl J Med. 1974 May 23;290(21):1177–1184. doi: 10.1056/NEJM197405232902106. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Kasper D. L., Seiler M. W. Immunochemical characterization of the outer membrane complex of Bacteroides fragilis subspecies fragilis. J Infect Dis. 1975 Oct;132(4):440–450. doi: 10.1093/infdis/132.4.440. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Linder L., Lindquist L., Söder P. O., Holme T. Estimation of cell lysis. Determination of aminopeptidase in extracts of Streptococcus mitis, ATCC 903. Acta Pathol Microbiol Scand B Microbiol Immunol. 1974 Oct;82B(5):602–607. doi: 10.1111/j.1699-0463.1974.tb00226.x. [DOI] [PubMed] [Google Scholar]
  13. Mathew A., Harris A. M., Marshall M. J., Ross G. W. The use of analytical isoelectric focusing for detection and identification of beta-lactamases. J Gen Microbiol. 1975 May;88(1):169–178. doi: 10.1099/00221287-88-1-169. [DOI] [PubMed] [Google Scholar]
  14. Neu H. C. The 5'-nucleotidase of Escherichia coli. I. Purification and properties. J Biol Chem. 1967 Sep 10;242(17):3896–3904. [PubMed] [Google Scholar]
  15. Neu H. C. The cyclic phosphodiesterases (3'-nucleotidases) of the enterobacteriaceae. Biochemistry. 1968 Oct;7(10):3774–3780. doi: 10.1021/bi00850a060. [DOI] [PubMed] [Google Scholar]
  16. Neu H. C. The surface localization of penicillinases in Escherichia coli and Salmonella typhimurium. Biochem Biophys Res Commun. 1968 Jul 26;32(2):258–263. doi: 10.1016/0006-291x(68)90378-1. [DOI] [PubMed] [Google Scholar]
  17. Neu H. C., Winshell E. B. Relation of beta-lactamase activity and cellular location to resistance of Enterobacter to penicillins and cephalosporins. Antimicrob Agents Chemother. 1972 Feb;1(2):107–111. doi: 10.1128/aac.1.2.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nord C. E., Möllby R., Smyth C., Wadström T. Formation of phospholipase C and theta-haemolysin in pre-reduced media in batch anc continuous culture of Clostridium perfringens type A. J Gen Microbiol. 1974 Sep;84(1):117–127. doi: 10.1099/00221287-84-1-117. [DOI] [PubMed] [Google Scholar]
  19. Nord C. E., Wadström T. Formation of -L- and -D-fucosidase in cultures of Streptococcus mitis. Med Microbiol Immunol. 1972;158(2):95–103. doi: 10.1007/BF02120474. [DOI] [PubMed] [Google Scholar]
  20. O'Callaghan C. H., Morris A., Kirby S. M., Shingler A. H. Novel method for detection of beta-lactamases by using a chromogenic cephalosporin substrate. Antimicrob Agents Chemother. 1972 Apr;1(4):283–288. doi: 10.1128/aac.1.4.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. O'Callaghan C., Morris A. Inhibition of beta-lactamases by beta-lactam antibiotics. Antimicrob Agents Chemother. 1972 Dec;2(6):442–448. doi: 10.1128/aac.2.6.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pinkus G., Veo G., Braude A. I. Bacteroides penicillinase. J Bacteriol. 1968 Oct;96(4):1437–1438. doi: 10.1128/jb.96.4.1437-1438.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Rubin F. A., Smith D. H. Characterization of R factor beta-lactamases by the acidimetric method. Antimicrob Agents Chemother. 1973 Jan;3(1):68–73. doi: 10.1128/aac.3.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Smith J. T., Wyatt J. M. Relation of R factor and chromosomal beta-lactamase with the periplasmic space. J Bacteriol. 1974 Mar;117(3):931–939. doi: 10.1128/jb.117.3.931-939.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Söderholm J., Allestam P., Wadström T. A rapid method for isoelectric focusing in polyacrylamide gel. FEBS Lett. 1972 Jul 15;24(1):89–92. doi: 10.1016/0014-5793(72)80833-0. [DOI] [PubMed] [Google Scholar]
  27. Wahren A., Holme T. Amino acid and peptide requirement of Fusiformis necrophorus. J Bacteriol. 1973 Oct;116(1):279–284. doi: 10.1128/jb.116.1.279-284.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wahren A., Holme T. Growth of Bacteroidaceae in stirred fermentors. Appl Microbiol. 1969 Aug;18(2):235–239. doi: 10.1128/am.18.2.235-239.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES