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. 1997 Apr;41(4):785–790. doi: 10.1128/aac.41.4.785

OXA-15, an extended-spectrum variant of OXA-2 beta-lactamase, isolated from a Pseudomonas aeruginosa strain.

F Danel 1, L M Hall 1, D Gur 1, D M Livermore 1
PMCID: PMC163795  PMID: 9087490

Abstract

Pseudomonas aeruginosa AH, isolated in Ankara, Turkey, was highly resistant to ceftazidime (MIC, 128 microg/ml) and produced a beta-lactamase that gave a doublet of bands at pIs 8.7 and 8.9. beta-Lactamase production was transferable to P. aeruginosa PU21 by conjugation and was determined by a ca. 450-kb plasmid, pMLH54. The transconjugant and Escherichia coli transformed with the cloned gene showed increased resistance to ceftazidime (especially) and to cefpirome, ceftazidime, ceftriaxone, moxalactam, and aztreonam, but not to carbapenems. Resistance was not reversed by clavulanic acid or tazobactam. Sequencing revealed that the beta-lactamase responsible for this resistance was identical to OXA-2 except that glycine replaced aspartate at position 150. Compared to OXA-2, the new enzyme, named OXA-15, had greater cephalosporinase activity, with increased relative hydrolysis rates for cephaloridine and cephalothin and, most dramatically, for ceftazidime. Cefotaxime and carbapenems remained stable to hydrolysis. Thus, as in the TEM, SHV, and OXA-10 (PSE-2) beta-lactamase families, a minor sequence change in OXA-2 gave a major extension of cephalosporinase activity and contingent resistance. The gene encoding the new beta-lactamase, bla(OXA-15), lay close to the highly conserved 3' end of an integron and had flanking sequences typical of an integron-associated gene cassette. Restriction mapping and partial sequence data indicated that pMLH54 carries an integron with three putative gene cassettes: bla(OXA-15) itself, aadB [coding aminoglycoside nucleotidyltransferase (2")-1a], and an uncharacterized cassette.

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Selected References

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  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Bush K., Jacoby G. A., Medeiros A. A. A functional classification scheme for beta-lactamases and its correlation with molecular structure. Antimicrob Agents Chemother. 1995 Jun;39(6):1211–1233. doi: 10.1128/aac.39.6.1211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cameron F. H., Groot Obbink D. J., Ackerman V. P., Hall R. M. Nucleotide sequence of the AAD(2'') aminoglycoside adenylyltransferase determinant aadB. Evolutionary relationship of this region with those surrounding aadA in R538-1 and dhfrII in R388. Nucleic Acids Res. 1986 Nov 11;14(21):8625–8635. doi: 10.1093/nar/14.21.8625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen H. Y., Yuan M., Livermore D. M. Mechanisms of resistance to beta-lactam antibiotics amongst Pseudomonas aeruginosa isolates collected in the UK in 1993. J Med Microbiol. 1995 Oct;43(4):300–309. doi: 10.1099/00222615-43-4-300. [DOI] [PubMed] [Google Scholar]
  5. Chou P. Y., Fasman G. D. Prediction of beta-turns. Biophys J. 1979 Jun;26(3):367–383. doi: 10.1016/S0006-3495(79)85259-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dale J. W., Godwin D., Mossakowska D., Stephenson P., Wall S. Sequence of the OXA2 beta-lactamase: comparison with other penicillin-reactive enzymes. FEBS Lett. 1985 Oct 21;191(1):39–44. doi: 10.1016/0014-5793(85)80989-3. [DOI] [PubMed] [Google Scholar]
  7. Dale J. W., Smith J. T. The purification and properties of the -lactamase specified by the resistance factor R-1818 in Escherichia coli and Proteus mirabilis. Biochem J. 1971 Jul;123(4):493–500. doi: 10.1042/bj1230493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Danel F., Hall L. M., Gur D., Akalin H. E., Livermore D. M. Transferable production of PER-1 beta-lactamase in Pseudomonas aeruginosa. J Antimicrob Chemother. 1995 Feb;35(2):281–294. doi: 10.1093/jac/35.2.281. [DOI] [PubMed] [Google Scholar]
  9. Danel F., Hall L. M., Gur D., Livermore D. M. OXA-14, another extended-spectrum variant of OXA-10 (PSE-2) beta-lactamase from Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1995 Aug;39(8):1881–1884. doi: 10.1128/aac.39.8.1881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Deléage G., Roux B. An algorithm for protein secondary structure prediction based on class prediction. Protein Eng. 1987 Aug-Sep;1(4):289–294. doi: 10.1093/protein/1.4.289. [DOI] [PubMed] [Google Scholar]
  11. Du Bois S. K., Marriott M. S., Amyes S. G. TEM- and SHV-derived extended-spectrum beta-lactamases: relationship between selection, structure and function. J Antimicrob Chemother. 1995 Jan;35(1):7–22. doi: 10.1093/jac/35.1.7. [DOI] [PubMed] [Google Scholar]
  12. Hall L. M., Livermore D. M., Gur D., Akova M., Akalin H. E. OXA-11, an extended-spectrum variant of OXA-10 (PSE-2) beta-lactamase from Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1993 Aug;37(8):1637–1644. doi: 10.1128/aac.37.8.1637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hall R. M., Brookes D. E., Stokes H. W. Site-specific insertion of genes into integrons: role of the 59-base element and determination of the recombination cross-over point. Mol Microbiol. 1991 Aug;5(8):1941–1959. doi: 10.1111/j.1365-2958.1991.tb00817.x. [DOI] [PubMed] [Google Scholar]
  14. Hall R. M., Brown H. J., Brookes D. E., Stokes H. W. Integrons found in different locations have identical 5' ends but variable 3' ends. J Bacteriol. 1994 Oct;176(20):6286–6294. doi: 10.1128/jb.176.20.6286-6294.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hall R. M., Collis C. M. Mobile gene cassettes and integrons: capture and spread of genes by site-specific recombination. Mol Microbiol. 1995 Feb;15(4):593–600. doi: 10.1111/j.1365-2958.1995.tb02368.x. [DOI] [PubMed] [Google Scholar]
  16. Hansen J. B., Olsen R. H. Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5. J Bacteriol. 1978 Jul;135(1):227–238. doi: 10.1128/jb.135.1.227-238.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Huovinen P., Huovinen S., Jacoby G. A. Sequence of PSE-2 beta-lactamase. Antimicrob Agents Chemother. 1988 Jan;32(1):134–136. doi: 10.1128/aac.32.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jacoby G. A., Medeiros A. A. More extended-spectrum beta-lactamases. Antimicrob Agents Chemother. 1991 Sep;35(9):1697–1704. doi: 10.1128/aac.35.9.1697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jacoby G. A. Properties of R plasmids determining gentamicin resistance by acetylation in Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1974 Sep;6(3):239–252. doi: 10.1128/aac.6.3.239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Joris B., Ledent P., Dideberg O., Fonzé E., Lamotte-Brasseur J., Kelly J. A., Ghuysen J. M., Frère J. M. Comparison of the sequences of class A beta-lactamases and of the secondary structure elements of penicillin-recognizing proteins. Antimicrob Agents Chemother. 1991 Nov;35(11):2294–2301. doi: 10.1128/aac.35.11.2294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kado C. I., Liu S. T. Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol. 1981 Mar;145(3):1365–1373. doi: 10.1128/jb.145.3.1365-1373.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ledent P., Frère J. M. Substrate-induced inactivation of the OXA2 beta-lactamase. Biochem J. 1993 Nov 1;295(Pt 3):871–878. doi: 10.1042/bj2950871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ledent P., Raquet X., Joris B., Van Beeumen J., Frère J. M. A comparative study of class-D beta-lactamases. Biochem J. 1993 Jun 1;292(Pt 2):555–562. doi: 10.1042/bj2920555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Li X. Z., Ma D., Livermore D. M., Nikaido H. Role of efflux pump(s) in intrinsic resistance of Pseudomonas aeruginosa: active efflux as a contributing factor to beta-lactam resistance. Antimicrob Agents Chemother. 1994 Aug;38(8):1742–1752. doi: 10.1128/aac.38.8.1742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Liu P. Y., Gur D., Hall L. M., Livermore D. M. Survey of the prevalence of beta-lactamases amongst 1000 gram-negative bacilli isolated consecutively at the Royal London Hospital. J Antimicrob Chemother. 1992 Oct;30(4):429–447. doi: 10.1093/jac/30.4.429. [DOI] [PubMed] [Google Scholar]
  26. Livermore D. M. beta-Lactamases in laboratory and clinical resistance. Clin Microbiol Rev. 1995 Oct;8(4):557–584. doi: 10.1128/cmr.8.4.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lévesque C., Piché L., Larose C., Roy P. H. PCR mapping of integrons reveals several novel combinations of resistance genes. Antimicrob Agents Chemother. 1995 Jan;39(1):185–191. doi: 10.1128/aac.39.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Mugnier P., Dubrous P., Casin I., Arlet G., Collatz E. A TEM-derived extended-spectrum beta-lactamase in Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1996 Nov;40(11):2488–2493. doi: 10.1128/aac.40.11.2488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nordmann P., Naas T. Sequence analysis of PER-1 extended-spectrum beta-lactamase from Pseudomonas aeruginosa and comparison with class A beta-lactamases. Antimicrob Agents Chemother. 1994 Jan;38(1):104–114. doi: 10.1128/aac.38.1.104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nordmann P., Ronco E., Naas T., Duport C., Michel-Briand Y., Labia R. Characterization of a novel extended-spectrum beta-lactamase from Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1993 May;37(5):962–969. doi: 10.1128/aac.37.5.962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ouellette M., Bissonnette L., Roy P. H. Precise insertion of antibiotic resistance determinants into Tn21-like transposons: nucleotide sequence of the OXA-1 beta-lactamase gene. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7378–7382. doi: 10.1073/pnas.84.21.7378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Paulsen I. T., Littlejohn T. G., Rådström P., Sundström L., Sköld O., Swedberg G., Skurray R. A. The 3' conserved segment of integrons contains a gene associated with multidrug resistance to antiseptics and disinfectants. Antimicrob Agents Chemother. 1993 Apr;37(4):761–768. doi: 10.1128/aac.37.4.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rasmussen B. A., Keeney D., Yang Y., Bush K. Cloning and expression of a cloxacillin-hydrolyzing enzyme and a cephalosporinase from Aeromonas sobria AER 14M in Escherichia coli: requirement for an E. coli chromosomal mutation for efficient expression of the class D enzyme. Antimicrob Agents Chemother. 1994 Sep;38(9):2078–2085. doi: 10.1128/aac.38.9.2078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sanschagrin F., Couture F., Levesque R. C. Primary structure of OXA-3 and phylogeny of oxacillin-hydrolyzing class D beta-lactamases. Antimicrob Agents Chemother. 1995 Apr;39(4):887–893. doi: 10.1128/aac.39.4.887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Shaw K. J., Rather P. N., Hare R. S., Miller G. H. Molecular genetics of aminoglycoside resistance genes and familial relationships of the aminoglycoside-modifying enzymes. Microbiol Rev. 1993 Mar;57(1):138–163. doi: 10.1128/mr.57.1.138-163.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Smith J. T. R-factor gene expression gram-negative bacteria. J Gen Microbiol. 1969 Jan;55(1):109–120. doi: 10.1099/00221287-55-1-109. [DOI] [PubMed] [Google Scholar]
  38. Stokes H. W., Hall R. M. A novel family of potentially mobile DNA elements encoding site-specific gene-integration functions: integrons. Mol Microbiol. 1989 Dec;3(12):1669–1683. doi: 10.1111/j.1365-2958.1989.tb00153.x. [DOI] [PubMed] [Google Scholar]
  39. Stokes H. W., Hall R. M. The integron In1 in plasmid R46 includes two copies of the oxa2 gene cassette. Plasmid. 1992 Nov;28(3):225–234. doi: 10.1016/0147-619x(92)90054-e. [DOI] [PubMed] [Google Scholar]
  40. Vivian A. Plasmid expansion? Microbiology. 1994 Feb;140(Pt 2):213–214. doi: 10.1099/13500872-140-2-213-a. [DOI] [PubMed] [Google Scholar]
  41. Waley S. G. The kinetics of substrate-induced inactivation. Biochem J. 1991 Oct 1;279(Pt 1):87–94. doi: 10.1042/bj2790087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Walsh T. R., Hall L., MacGowan A. P., Bennett P. M. Sequence analysis of two chromosomally mediated inducible beta-lactamases from Aeromonas sobria, strain 163a, one a class D penicillinase, the other an AmpC cephalosporinase. J Antimicrob Chemother. 1995 Jul;36(1):41–52. doi: 10.1093/jac/36.1.41. [DOI] [PubMed] [Google Scholar]
  43. Watanabe M., Iyobe S., Inoue M., Mitsuhashi S. Transferable imipenem resistance in Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1991 Jan;35(1):147–151. doi: 10.1128/aac.35.1.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  45. Zhu Y., Englebert S., Joris B., Ghuysen J. M., Kobayashi T., Lampen J. O. Structure, function, and fate of the BlaR signal transducer involved in induction of beta-lactamase in Bacillus licheniformis. J Bacteriol. 1992 Oct;174(19):6171–6178. doi: 10.1128/jb.174.19.6171-6178.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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