Article Figures & Data
Figures
- Fig. 1.
(A) Agarose (0.7%) electrophoresis of plasmid DNA from ESBL-producing E. coli transconjugants and from IRT-producing P. mirabilis. (B) Hybridization with TEM-specific DNA probes. Lanes: 1, plasmid RSa (39 kb); 2, plasmid TP114 (61 kb); 3, pCFF04 (TEM-3; 85 kb); 4, pCFF74 (TEM-24; 85 kb); 5, pCFF14 (TEM-5; 180 kb); 6, pCFF134 (TEM-3); 7, TrCF669 (TEM-66); 8, TrCF39 (TEM-3); 9, TrCF249 (TEM-24); 10, CF449 (TEM-44); 11, CF659 (TEM-65).
Tables
- Table 1.
β-Lactam MICs for TEM-producingP. mirabilis strains
β-Lactam MIC (μg/ml) for the following strains (enzymes): CF19 (TEM-1) CF29 (TEM-2) CF579 (TEM-57) Amoxicillin 512 4,096 4,096 Amoxicillin + CAa 4 16 16 Ticarcillin 128 512 512 Ticarcillin + CA 2 2 2 Piperacillin 8 256 256 Piperacillin + TAZb 0.5 1 1 Cephalothin 8 8 8 - Table 2.
β-Lactam MICs for ESBL-producing P. mirabilis strains and their E. coli transconjugants
β-Lactams MIC (μg/ml) for the following strains (enzymes): P. mirabilis CF249 (TEM-24) E. coli TrCF249 (TEM-24) P. mirabilis CF39 (TEM-3) E. coliTrCF39 (TEM-3) P. mirabilis CF669 (TEM-66) E. coli TrCF669 (TEM-66) Cefotaxime 0.12 1 2 4 8 4 Cefotaxime + CAa 0.03 0.03 0.06 0.06 0.06 0.03 Ceftazidime 4 64 0.5 8 8 4 Ceftazidime + CA 0.12 0.25 0.12 0.25 0.12 0.12 Aztreonam 0.06 4 0.06 2 0.25 2 Aztreonam + CA 0.01 0.03 0.01 0.06 0.06 0.003 ↵a CA, clavulanic acid (2 μg/ml).
- Table 3.
β-Lactam MICs for IRT-producingP. mirabilis strains
β-Lactam MIC (μg/ml) for the following strains (enzymes): CF739 (TEM-73 [IRT-18]) CF749 (TEM-74 [IRT-19]) CF449 (TEM-44 [IRT-13]) CF659 (TEM-65 [IRT-16]) Amoxicillin 1,024 2,048 1,024 64 Amoxicillin + CAa 1,024 512 512 64 Ticarcillin 8 32 16 1 Ticarcillin + CA 4 4 2 1 Piperacillin 16 16 8 2 Piperacillin + TAZb 4 1 0.5 0.5 Cephalothin 2 4 4 2 - Table 4.
pIs and kinetic parameters inhibitors for TEM-type and IRT-type β-lactamases
Drug Kinetic parameters for the following strains (enzymes; pIs): CF19 (TEM-1; 5.4) CF29 (TEM-2; 5.6) CF579 (TEM-57; 5.2) CF739 (TEM-73 [IRT-18]; 5.2) CF749 (TEM-74 [IRT-19]; 5.2) CF449 (TEM-44 [IRT-13]; 5.4) CF659 (TEM-65 [IRT-16]; 5.4) Vmaxa Km(μM) Vmax Km(μM) Vmax Km(μM) Vmax Km(μM) Vmax Km(μM) Vmax Km(μM) Vmax Km(μM) Benzylpenicillin 100 25 100 20 100 40 100 334 100 330 100 375 100 350 Amoxicillin 77 26 95 25 73 38 207 315 140 400 145 360 150 400 Ticarcillin 10 10 10 9 10 15 2 810 2 765 2.3 460 2 600 Cephalothin 10 250 25 500 17 900 1.5 1,000 1 300 2 312 2 400 ↵a Vmax values are given as a percentage of the Vmax for benzylpenicillin, which was taken as 100%.
- Table 5.
IC50s of inhibitors for TEM-type and IRT-type β-lactamases
Strain (enzyme) IC50 (μM) Clavulanate Tazobactam CF19 (TEM-1) 0.18 0.05 CF29 (TEM-2) 0.09 0.04 CF579 (TEM-57) NDa ND CF739 (TEM-73 [IRT-18]) 31 12 CF749 (TEM-74 [IRT-19]) 9.4 2.9 CF449 (TEM-44 [IRT-13]) 10 3.0 CF65a [TEM-65 [IRT-16]) 9.0 3.0 ↵a ND, not done.
- Table 6.
pI and kinetic parameters for ESBL-type β-lactamases
Drug Kinetic parameters for the following strains (enzymes; pIs): CF249 (TEM-24; 6.5) CF39 (TEM-3; 6.3) CF669 (TEM-66; 6.0) Vmaxa Km(μM) Vmax Km(μM) Vmax Km(μM) Benzylpenicillin 100 5 100 4 100 5 Amoxicillin 62 43 72 5 70 5 Cephalothin 281 43 107 22 110 30 Cefotaxime 130 50 445 100 400 100 Ceftazidime 1,410 377 40 400 40 450 ↵a Vmax values are given as a percentage of the Vmax for benzylpenicillin, which was taken as 100%.
- Table 7.
Nucleotide and amino acid substitutions in theblaTEM genes fromP. mirabilis strains
β-Lactamase and strain or enzyme Mutation at the following nucleotide (substituted amino acid)a: 32b 226 (8) 263 (21) 317 (39) 346 (48) 436 (78) 477 (92) 512 (104) 604 (134) 682 (160) 692 (164) 911 (237) 914 (238) 917 (240) 925 (242) 929 (244) 990 (265) TEM-like TEM-1 (blaTEM-1A)c GCC-TA C (Phe) C (Leu) C (Gln) A (Glu) C (Gly) G (Gly) G (Glu) G (Ala) T (Thr) C (Arg) G (Ala) G (Gly) G (Glu) G (Gly) C (Arg) C (Thr) TEM-1 (blaTEM-1B)c GCC-TA T T T TEM-2 (blaTEM-2)c GCT-TA A (Lys) G T C A CF579 (TEM-57) GCTATA G T A (Asp) C A ESBL CF249 (TEM-24) GCT-TA A (Lys) G T A (Lys) C A (Ser) A (Thr) A (Lys) A CF39 (TEM-3) GCT-TA A (Lys) G T A (Lys) A (Ser) A CF669 (TEM-66) GCT-TA A (Lys) G T A (Asp) A (Lys) A (Ser) IRT-like CF449 (TEM-44 [IRT-13]) GCT-TA A (Lys) G T C A A (Ser) CF659 (TEM-65 [IRT-16]) GCT-TA A (Lys) G C A T (Cys) CF739 (TEM-73 [IRT-18]) GCT-TA T (Phe) C G T C A T (Cys) T (Met) CF749 (TEM-74 [IRT-19]) GCT-TA T (Phe) C G T C A A (Ser) T (Met)
