Characterization of cell-bound papain-soluble beta-lactamases in BRO-1 and BRO-2 producing strains ofMoraxella (Branhamella) catarrhalis andMoraxella nonliquefaciens

  • I. Eliasson
  • C. Kamme
  • M. Vang
  • S. G. Waley


InMoraxella (Branhamella) catarrhalis andMoraxella nonliquefaciens strains isolated from clinical specimens in the south of Sweden two variants of beta-lactamase were distinguished by isoelectric focusing (IEF). The BRO-1 (Ravasio type) enzyme was the most common inBranhamella catarrhalis, constituting about 90% of the beta-lactamase found in this species, while the BRO-2 enzyme (1908 type) was as common as BRO-1 inMoraxella nonliquefaciens. The determinants mediating the production of BRO-1 and BRO-2 were both transferable by conjugation. Cell-bound beta-lactamase from reference strains producing BRO-1 and BRO-2 could be solubilized by papain digestion. The isoelectric point of the solubilized enzymes differed distinctly between BRO-1 (pI 6.5) and BRO-2 (pI 6.9). The molecular species of BRO-1 and BRO-2 released by papain digestion were purified by affinity chromatography with phenylboronic acid agarose gel. They had identical molecular weights of approximately 28,000. Their kinetic constants were indistinguishable for a number of substrates and beta-lactamase inhibitors.


Molecular Weight Internal Medicine Agarose Isoelectric Point Affinity Chromatography 
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  1. 1.
    Malmvall BE, Brorsson JE, Johnsson J: In vitro sensitivity to penicillin V and beta-lactamase production ofBranhamella catarrhalis. Journal of Antimicrobial Chemotherapy 1977, 3: 374–375.Google Scholar
  2. 2.
    Percival A, Corkill JE, Rowlands J, Sykes RB: Pathogenicity of and β-lactamase production byBranhamella (Neisseria) catarrhalis. Lancet 1977: 1175.Google Scholar
  3. 3.
    Rosenthal SL, Freundlich LF, Gilardi GL, Clodomar FY: In vitro antibiotic sensitivity ofMoraxella species. Chemotherapy 1978, 24: 360–363.Google Scholar
  4. 4.
    Alvarez S, Jones M, Holtsclaw-Berk S, Guarderas J, Berk SL: In vitro susceptibility and β-lactamase production of 53 clinical isolates ofBranhamella catarrhalis. Antimicrobial Agents and Chemotherapy 1985, 27: 646–647.Google Scholar
  5. 5.
    Wallace RJ, Steingrube VA, Nash DR, Hollis DG, Flanagan C, Brown BA, Labidi A, Weaver RE: BRO β-lactamases ofBranhamella catarrhalis andMoraxella subgenusMoraxella, including evidence for chromosomal β-lactamase transfer by conjugation inB. catarrhalis, M. nonliquefaciens andM. lacunata. Antimicrobial Agents and Chemotherapy 1989, 33: 1845–1854.Google Scholar
  6. 6.
    McLeod DT, Ahmad F, Croughan MJ, Calder MA: Bronchopulmonary infection due toB. catarrhalis: clinical features and therapeutic response. Drugs 1986, 31, Supplement 3: 109–112.Google Scholar
  7. 7.
    Philippon A, Riou JY, Guibourdenche M, Sotolongo F: Detection, distribution and inhibition ofBranhamella catarrhalis β-lactamases. Drugs 1986, 31, Supplement 3: 64–69.Google Scholar
  8. 8.
    Saito A, Yamaguchi K, Shigeno Y, Kohno S, Shigeno H, Kusano N, Dotsu Y, Hara K: Clinical and bacteriological evaluation ofBranhamella catarrhalis in respiratory infections. Drugs 1986, 31, Supplement 3: 87–92.Google Scholar
  9. 9.
    Eliasson I, Kamme C: Upper respiratory tract infections. Ecological and therapeutic aspects of beta-lactamase production with special reference toBranhamella catarrhalis. Drugs 1986, 31, Supplement 3: 116–121.Google Scholar
  10. 10.
    Eliasson I, Holst E, Mölstad S, Kamme C: Emergence and persistence of β-lactamase-producing bacteria in the upper respiratory tract in children treated with β-lactam antibiotics. American Journal of Medicine 1990, 88, Supplement 5A: 51–55.Google Scholar
  11. 11.
    Farmer T, Reading C: Beta-lactamases ofBranhamella catarrhalis and their inhibition by clavulanic acid. Antimicrobial Agents and Chemotherapy 1982, 21: 506–508.Google Scholar
  12. 12.
    Kamme C, Eliasson I, Kahl-Knutson B, Wang M: Plasmid-mediated beta-lactamase inBranhamella catarrhalis. Drugs 1986, 31, Supplement 3: 55–63.Google Scholar
  13. 13.
    Eliasson I, Kamme C, Prellner K: Beta-lactamase production in the upper respiratory tract flora. European Journal of Clinical Microbiology 1986, 5: 507–512.Google Scholar
  14. 14.
    Kamme C, Vang M, Ståhl S: Intrageneric and intergeneric transfer ofBranhamella catarrhalis beta-lactamase production. Scandinavian Journal of Infectious Diseases 1984, 16: 153–155.Google Scholar
  15. 15.
    Nash DR, Wallace RJ, Steingrube VA, Shurin PA: Isoelectric focusing of β-lactamases from sputum and middle ear isolates ofBranhamella catarrhalis recovered in the United States. Drugs 1986, 31, Supplement 3: 48–54.Google Scholar
  16. 16.
    Simpson IN, Plested SJ: The origin and properties of β-lactamase satellite bands seen in isoelectric focusing. Journal of Antimicrobial Chemotherapy 1983, 12: 127–131.Google Scholar
  17. 17.
    Stobberingh EE, Davies BI, van Boven CPA:Branhamella catarrhalis: antibiotic sensitivities and betalactamases. Journal of Antimicrobial Chemotherapy 1984, 13: 55–64.Google Scholar
  18. 18.
    Thorne GM, Farrar WE: Transfer of ampicillin resistance between strains ofHaemophilus influenzae type B. Journal of Infectious Diseases 1975, 132: 276–281.Google Scholar
  19. 19.
    Smith MD, Guild WR: Improved method for conjugative transfer by filter mating ofStreptococcus pneumoniae. Journal of Bacteriology 1980, 144: 457–459.Google Scholar
  20. 20.
    Eliasson I, Kamme C: Characterization of the plasmid-mediated beta-lactamase inBranhamella catarrhalis with special reference to substrate affinity. Journal of Antimicrobial Chemotherapy 1985, 15: 139–149.Google Scholar
  21. 21.
    Bhaduri S, Demchick PH: Simple and rapid method for disruption of bacteria for protein studies. Applied Environmental Microbiology 1983, 46: 941–943.Google Scholar
  22. 22.
    Cartwright SJ, Waley SG: Purification of β-lactamases by affinity chromatography on phenylboronic acid-agarose. Biochemical Journal 1984, 221: 505–512.Google Scholar
  23. 23.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the folin phenol reagent. Journal of Biological Chemistry 1951, 193: 265–275.Google Scholar
  24. 24.
    Neville DM: Molecular weight determination of protein-dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer system. Journal of Biological Chemistry 1971, 246: 6328–6334.Google Scholar
  25. 25.
    Wharton W, Szawelski RJ: Half-time analysis of the integrated Michaelis equation. Biochemical Journal 1982, 203: 351–360.Google Scholar
  26. 26.
    Waley SG: A quick method for the determination of inhibition constants. Biochemical Journal 1982, 205: 631–633.Google Scholar
  27. 27.
    De Meester, F, Frère JM, Waley SG, Cartwright SJ, Virden R, Lindberg F: 6-β-iodopenicillanate as a probe for the classification of β-lactamases. Biochemical Journal 1986, 239: 575–580.Google Scholar
  28. 28.
    Waley SG: Kinetics of suicide substrates. Biochemical Journal 1985, 227: 843–849.Google Scholar
  29. 29.
    Beaulieu D, Ouellette M, Bergeron MG, Roy PH: Characterization of a plasmid isolated fromBranhamella catarrhalis and detection of plasmid sequences within the genome of aB. catarrhalis strain. Plasmid 1988, 20: 158–162.Google Scholar
  30. 30.
    Biörck L, Kronvall G: Purification and some properties of streptococcal protein G, a novel IgG-binding reagent. Journal of Immunology 1984, 133: 969–974.Google Scholar
  31. 31.
    Farmer T, Reading C: Inhibition of the β-lactamases ofBranhamella catarrhalis by clavulanic acid and other inhibitors. Drugs 1986, 31, Supplement 3: 70–78.Google Scholar
  32. 32.
    Beesley T, Gascoyne N, Knott-Hunziker V, Petursson S, Waley SG, Jaurin B, Grundström T: The inhibition of class C β-lactamases by boronic acids. Biochemical Journal 1983, 209: 229–233.Google Scholar
  33. 33.
    Yokota E, Fujii T, Sato K, Inoue M, Mitsuhashi S: Purification and properties of a β-lactamase produced byBranhamella catarrhalis. Antimicrobial Agents and Chemotherapy 1986, 29: 696–698.Google Scholar
  34. 34.
    Luman I, Wilson RW, Wallace RJ, Nash DR: Disk diffusion susceptibility ofBranhamella catarrhalis and relationship of β-lactam zone size to β-lactamase production. Antimicrobial Agents and Chemotherapy 1986, 30: 774–776.Google Scholar
  35. 35.
    Hoi-Dang Van AB, Bourguenec CBL, Barthelemy M, Labia R: Novel β-lactamase fromBranhamella catarrhalis. Annals of Microbiology 1978, 129B: 397–406.Google Scholar
  36. 36.
    Bush K: Characterization of β-lactamases. Antimicrobial Agents and Chemotherapy 1989, 33: 259–263.Google Scholar
  37. 37.
    Bush K: Classification of β-lactamases: groups 2c, 2d, 2e, 3 and 4. Antimicrobial Agents and Chemotherapy 1989, 33: 271–276.Google Scholar

Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1992

Authors and Affiliations

  • I. Eliasson
    • 1
  • C. Kamme
    • 1
  • M. Vang
    • 1
  • S. G. Waley
    • 2
  1. 1.Department of Medical MicrobiologyUniversity of LundLundSweden
  2. 2.Sir William Dunn School of PathologyUniversity of OxfordOxfordUK

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