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Mechanisms of Resistance

Characterization of TEM-56, a Novel β-Lactamase Produced by a Klebsiella pneumoniae Clinical Isolate

Catherine Neuwirth, Roger Labia, Eliane Siebor, Andre Pechinot, Stephanie Madec, El Bachir Chaibi, Antoine Kazmierczak
Catherine Neuwirth
Laboratoire de Bactériologie, Hôpital Universitaire du Bocage, B.P. 1542, 21034 Dijon Cedex, and
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Roger Labia
Centre National de la Recherche Scientifique, UMR175, 6 rue de l'Université, 29000 Quimper, France
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Eliane Siebor
Laboratoire de Bactériologie, Hôpital Universitaire du Bocage, B.P. 1542, 21034 Dijon Cedex, and
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Andre Pechinot
Laboratoire de Bactériologie, Hôpital Universitaire du Bocage, B.P. 1542, 21034 Dijon Cedex, and
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Stephanie Madec
Centre National de la Recherche Scientifique, UMR175, 6 rue de l'Université, 29000 Quimper, France
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El Bachir Chaibi
Centre National de la Recherche Scientifique, UMR175, 6 rue de l'Université, 29000 Quimper, France
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Antoine Kazmierczak
Laboratoire de Bactériologie, Hôpital Universitaire du Bocage, B.P. 1542, 21034 Dijon Cedex, and
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DOI: 10.1128/AAC.44.2.453-455.2000
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Tables

  • Table 1.

    MICs for K. pneumoniae clinical isolates Kp 395 and Kp 377 and E. coli strains producing TEM-18, TEM-2, or TEM-3

    Strain or β-lactamase producedMIC (μg/ml)a
    AMXTICPIPCFTFOXATMCTXCAZ
    K. pneumoniae
     Kp 395 (TEM-56)>8,192 (1,024)>8,192 (1,024)>2,048 (16)5124 (4)1 (0.25)0.25 (≤0.06)4 (2)
     Kp 377 (TEM-21)>8,192 (32)>8,192 (16)1,024 (4)2564 (4)16 (0.12)128 (≤0.06)32 (1)
    E. coli
     TEM-188,192 (512)1,024 (64)512 (16)168 (8)2 (1)0.25 (0.12)4 (2)
     TEM-38,192 (16)8,192 (32)256 (2)1284 (4)8 (0.5)64 (0.12)32 (0.5)
     TEM-24,096 (16)4,096 (32)512 (2)322 (2)≤0.06 (≤0.06)≤0.06 (≤0.06)0.5 (0.25)
    • ↵a AMX, amoxicillin; TIC, ticarcillin; PIP, piperacillin; CFT, cephalothin; FOX, cefoxitin; ATM, aztreonam; CTX, cefotaxime; CAZ, ceftazidime. The values in parentheses are MICs in the presence of 2 μg/ml of clavulanic acid.

  • Table 2.

    Nucleotide substitutions inblaTEM-2, blaTEM-21, and blaTEM-56 genes

    Region and nucleotide positionaNucleotide in:
    blaTEM-2blaTEM-21blaTEM-56
    Promoter region
     32*bTTT
     175*AAA
    Coding region
     226*CCC
     317*A (Lys-39c)AA
     346*GGG
     436*TTT
     512G (Glu-104)A (Lysd)A (Lys)
     604*GGG
     660A (His-153)G (Arg)G (Arg)
     682*CCC
     914G (Gly-238)A (Ser)G
     925*AAA
    • ↵a Nucleotide numbering is according to that given by Sutcliffe (23) and amino acid numbering is according to Ambler et al. (1).

    • ↵b Nucleotides marked with an asterisk are involved in the differences between blaTEM-2,blaTEM-1A, and blaTEM-1Bgenes (9, 10, 23).

    • ↵c Gln for TEM-1.

    • ↵d For TEM-21 and TEM-56, there are indicated amino acids differing from TEM-2.

  • Table 3.

    Comparative kinetic parameters of β-lactamase TEM-2 and its mutants

    DrugKinetic parameters ofa:
    TEM-2TEM-18TEM-56TEM-3TEM-21
    kcatKmkcat/KmkcatKmkcat/KmkcatKmkcat/KmkcatKmkcat/KmkcatKmkcat/Km
    Benzylpenicillin1,50018831,40024581,40022634567.55095.5
    Amoxicillin1,20020601,3004131.71,33040333156.243202.15
    Ticarcillin1951216.2280647300743924.5732.3
    Piperacillin1,2754031.81,36040341,35038357212646123.8
    Cephalothin3003500.862251351.72501401.848232.118550.33
    Cephaloridine8258001.01,3601,2001.11,3501,2001.143301.41201201.0
    Cefoperazone7002303.05501703.25501803.028201.419300.63
    Cefuroxime31,000<0.01104800.0284700.0228900.3118850.21
    Ceftriaxone<1ND<0.01218000.03168400.0281801532100.25
    Cefotaxime1.5ND<0.01351,6600.02281,7000.0220210021204000.3
    Ceftazidime<1ND<0.01<1ND<0.01<1ND<0.01182000.14ND<0.01
    • ↵a kcat values are given in per second units, Km values are given in micromolar units, and kcat/Km values are given in per micromole per second units. The values given forkcat and Km were computed on the basis of at least three independent experiments. ND, not detected (high Km or Kivalues).

  • Table 4.

    50% inhibitory concentrations (IC50s) of β-lactamase inhibitors for β-lactamase TEM-2 and its mutants

    β-LactamaseIC50 (μM) of:
    Clavulanic acidSulbactamTazobactam
    TEM-20.1680.1
    TEM-180.0074.20.1
    TEM-560.0084.30.1
    TEM-30.010.40.01
    TEM-210.0120.07
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Characterization of TEM-56, a Novel β-Lactamase Produced by a Klebsiella pneumoniae Clinical Isolate
Catherine Neuwirth, Roger Labia, Eliane Siebor, Andre Pechinot, Stephanie Madec, El Bachir Chaibi, Antoine Kazmierczak
Antimicrobial Agents and Chemotherapy Feb 2000, 44 (2) 453-455; DOI: 10.1128/AAC.44.2.453-455.2000

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Characterization of TEM-56, a Novel β-Lactamase Produced by a Klebsiella pneumoniae Clinical Isolate
Catherine Neuwirth, Roger Labia, Eliane Siebor, Andre Pechinot, Stephanie Madec, El Bachir Chaibi, Antoine Kazmierczak
Antimicrobial Agents and Chemotherapy Feb 2000, 44 (2) 453-455; DOI: 10.1128/AAC.44.2.453-455.2000
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KEYWORDS

Klebsiella pneumoniae
beta-lactamases

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