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Mechanisms of Resistance

β-Lactamases of Kluyvera ascorbata, Probable Progenitors of Some Plasmid-Encoded CTX-M Types

Christel Humeniuk, Guillaume Arlet, Valerie Gautier, Patrick Grimont, Roger Labia, Alain Philippon
Christel Humeniuk
1Service de Bactériologie, CHU Cochin
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Guillaume Arlet
2Service de Bactériologie, Hôpital Tenon, UFR Saint-Antoine
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Valerie Gautier
2Service de Bactériologie, Hôpital Tenon, UFR Saint-Antoine
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Patrick Grimont
3Institut Pasteur, INSERM U389, Paris
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Roger Labia
4Université de Brest, CNRS Unité FRE 2125, Quimper, France
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Alain Philippon
1Service de Bactériologie, CHU Cochin
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  • For correspondence: alain.philippon@cch.ap-hop-paris.fr
DOI: 10.1128/AAC.46.9.3045-3049.2002
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    FIG. 1.

    Genetic environment of blaKLUA-1 gene encoding the β-lactamase produced by K. ascorbata strain CIP 82.95T. The regions surrounding the chromosomal blaKLUA-1 gene and the sequences adjacent to the plasmidic genes encoding β-lactamases CTX-M-2, CTX-M-4, CTX-M-5, CTX-M-6, CTX-M-7, and Toho-1 (2, 10, 11, 23) were compared. The RTR sequence and tnpA correspond to the right terminal repeat and transposase A, respectively, which are typical of ISEcp1.

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  • TABLE 1.

    MICs of 12 β-lactams for the 12 clinical K. ascorbata isolates, recipient strain E. coli XL-1, and the E. coli XL-1 transformant producing KLUA-1

    β-Lactam(s)aNo. of isolates for which MIC (μg/ml) was:MIC (μg/ml) for E. coli XL-1 recipient
    <0.030.060.1250.250.51248163264128R+R−
    Amoxicillin11442>1284
    Amoxicillin + CA3b34211
    Ticarcillin2343>1280.5
    Ticarcillin + CA5b24140.125
    Piperacillin14431281
    Piperacillin + TZ4b331120.12
    Cephalothin273>1280.5
    Cefuroxime165641
    Cefoxitin144321
    Cefotaxime372128<0.03
    Ceftriaxone26414<0.03
    Ceftazidime65140.06
    Cefepime9214<0.03
    Aztreonam46280.12
    Imipenem1650.25<0.03
    • ↵ a CA, 2 μg of clavulanic acid per ml; TZ, 4 μg of tazobactam per ml.

    • ↵ b MIC, <0.125 μg/ml.

    • c R−, E. coli recipient; R+, E. coli with recombinant plasmid producing KLUA-1.

  • TABLE 2.

    Kinetic constants (Vmax, Km) of β-lactamase produced by K. ascorbata strain CIP 82. 95Ta

    SubstrateVmax (relative)Km (μM)Vmax/Km (relative)
    Benzylpenicillin10017.5100
    Ticarcillin7.51013
    Cephalothin1,100141136
    Cefuroxime803540
    Cefotaxime1103755.1
    Ceftazidime<0.150<0.04
    Aztreonam0.31050.06
    Cefoxitin<0.1110<0.02
    • ↵ a V max was expressed relative to the Vmax for benzylpenicillin (which was set at 100%).

  • TABLE 3.

    Genotypic characterization of β-lactamase contents of K. ascorbata strains and β-lactamase pIs

    StrainpIResult of PCR with primer(s):GeneGenbank accession no.
    MA1-MA2 (208-751 bp; 543 bp)MA3 (1-430 bp; 429 bp)MA4 (461-883 bp; 422 bp)
    CIP 82.95T8.0+++ bla KLUA-1 AJ272538
    IP 15.798.4+++ bla KLUA-2 AJ251722
    IP 5.798.0+++ bla KLUA-3 AJ427461
    IP 8.798.0+++ bla KLUA-4 AJ427462
    IP 12.798.0+++ bla KLUA-5 AJ427463
    SL Ka996.9−−−
    IP 2.896.9−−−
    IP 3.898.0+++ bla KLUA-8 AJ427465
    TN Ka018.4+++ bla KLUA-9 AJ427466
    IP 43.508.0+++ bla KLUA-10 AJ427467
    IP 4450.948.4+++ bla KLUA-11 AJ427468
    IP 684.748.4+++ bla KLUA-12 AJ427469
  • TABLE 4.

    Identities of amino acid sequences of K. ascorbata β-lactamases and CTX-M, Toho, UOE β-lactamases

    Enzyme(s)Range % identity (reference) with clusters including:
    KLUA-1, -2, -3, -4, -5, -8, -9, -10, -11, and -12 (this work)CTX-M-1, -3, -10, -11, and -12 and UOE-1 (13, 18, 23a)CTX-M-2, -4, -5, -6, and -7 and Toho-1 (3, 10, 11, 23)CTX-M-9, -13, -14, and -15 and UOE-2 (21b)Toho-2 (15)CTX-M-8 (2)
    KLUA-1, -2, -3, -4, -5, -8, -9, -10, -11, and -1296-10077-8295-10078-8168-7182-84
    CTX-M-1, -3, -10, -11, and -12 and UOE-198-10077-8179-8070-7180-82
    CTX-M-2, -4, -5, -6, and -7 and Toho-197-10078-8168-7181-84
    CTX-M-9, -13, -14, and -15 and UOE-299-10087-8885-86
    Toho-210075
    CTX-M-8100
    • ↵ a Genbank references numbers AY005110 for CTX-M-11 and AY013478 for UOE-1.

    • ↵ b Genbank reference numbers AF252621 for CTX-M-13, AF252622 for CTX-M-14, AF252623 for CTX-M-15, and AF311345 for UOE-1.

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β-Lactamases of Kluyvera ascorbata, Probable Progenitors of Some Plasmid-Encoded CTX-M Types
Christel Humeniuk, Guillaume Arlet, Valerie Gautier, Patrick Grimont, Roger Labia, Alain Philippon
Antimicrobial Agents and Chemotherapy Sep 2002, 46 (9) 3045-3049; DOI: 10.1128/AAC.46.9.3045-3049.2002

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β-Lactamases of Kluyvera ascorbata, Probable Progenitors of Some Plasmid-Encoded CTX-M Types
Christel Humeniuk, Guillaume Arlet, Valerie Gautier, Patrick Grimont, Roger Labia, Alain Philippon
Antimicrobial Agents and Chemotherapy Sep 2002, 46 (9) 3045-3049; DOI: 10.1128/AAC.46.9.3045-3049.2002
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  • Top
  • Article
    • ABSTRACT
    • Bacterial strains.
    • Antimicrobial agents and susceptibility testing.
    • Kinetic and IEF analyses.
    • Genetic characterization of blaKLUA.
    • Genetic environment of blaKLUA-1.
    • Arguments for mobilization of chromosomal blaKLUA gene.
    • Nucleotide sequence accession number.
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

KEYWORDS

Enterobacteriaceae
plasmids
beta-lactamases

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