Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | plasmid pBC |
Protein Variants | Comment | Organism |
---|---|---|
T237A | the mutant enzyme exhibits elevated MICs for ampicillin, piperacillin, and the beta-lactambeta-lactamase inhibitor combinations. The T237A mutant enzyme demonstrates a lower kcat/Km for imipenem, cephalothin, and cefotaxime. The T237A mutant enzyme shows increased Kis for clavulanic acid, sulbactam, and tazobactam | plasmid pBC |
T237S | the mutant enzyme maintains MICs equivalent to those of the wild type against all of the beta-lactams tested, including carbapenems. The mutant enzyme displays substrate kinetics similar to those of the wild-type KPC-2 enzyme. The T237S mutant enzyme displays Kis similar to wilde-type enzyme | plasmid pBC |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
clavulanic acid | - |
plasmid pBC | |
Sulbactam | - |
plasmid pBC | |
tazobactam | - |
plasmid pBC |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.005 | - |
nitrocefin | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
0.006 | - |
piperacillin | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
0.006 | - |
nitrocefin | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
0.012 | - |
piperacillin | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
0.014 | - |
cephalothin | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
0.015 | - |
nitrocefin | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
0.015 | - |
piperacillin | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
0.018 | - |
cephalothin | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
0.019 | - |
Imipenem | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
0.021 | - |
Imipenem | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
0.03 | - |
cephalothin | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
0.031 | - |
Imipenem | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
0.168 | - |
cefotaxime | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
0.207 | - |
cefotaxime | pH 7.4, 22°C, wild-type enzyme | plasmid pBC |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
plasmid pBC | - |
vector SK(+)-blaKPC-2 | - |
Purification (Comment) | Organism |
---|---|
- |
plasmid pBC |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cefotaxime + H2O | - |
plasmid pBC | (2R)-5-[(acetyloxy)methyl]-2-[(R)-[[(2E)-2-(2-amino-1,3-thiazol-4-yl)-2-(methoxyimino)acetyl]amino](carboxy)methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
cephalothin + H2O | - |
plasmid pBC | (2R)-5-[(acetyloxy)methyl]-2-[(R)-carboxy[(thiophen-2-ylacetyl)amino]methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
imipenem + H2O | - |
plasmid pBC | (5R)-3-[[2-(carbonoimidoylamino)ethyl]sulfanyl]-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-4,5-dihydro-1H-pyrrole-2-carboxylic acid | - |
? | |
nitrocefin + H2O | - |
plasmid pBC | (2R)-2-[(R)-carboxy[(thiophen-2-ylacetyl)amino]methyl]-5-[(E)-2-(2,4-dinitrophenyl)ethenyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
piperacillin + H2O | - |
plasmid pBC | (2R,4S)-2-[(R)-carboxy[[(2R)-2-[[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonyl]amino]-2-phenylacetyl]amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? |
Synonyms | Comment | Organism |
---|---|---|
KPC-2 beta-lactamase | - |
plasmid pBC |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
6.8 | - |
Imipenem | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
19 | - |
Imipenem | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
21 | - |
Imipenem | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
26 | - |
cephalothin | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
42 | - |
piperacillin | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
42 | - |
piperacillin | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
43 | - |
cefotaxime | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
44 | - |
cephalothin | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
50 | - |
nitrocefin | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
51 | - |
piperacillin | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
52 | - |
nitrocefin | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
65 | - |
cephalothin | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
142 | - |
cefotaxime | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
191 | - |
nitrocefin | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.011 | - |
clavulanic acid | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
0.013 | - |
clavulanic acid | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
0.053 | - |
tazobactam | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
0.055 | - |
clavulanic acid | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
0.074 | - |
tazobactam | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
0.097 | - |
Sulbactam | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
0.167 | - |
Sulbactam | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
0.174 | - |
tazobactam | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
0.703 | - |
Sulbactam | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
220 | - |
Imipenem | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
260 | - |
cefotaxime | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
700 | - |
cefotaxime | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
900 | - |
cephalothin | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
1000 | - |
Imipenem | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
1000 | - |
Imipenem | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
3100 | - |
cephalothin | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
3400 | - |
piperacillin | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
3500 | - |
piperacillin | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC | |
3600 | - |
cephalothin | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
4000 | - |
piperacillin | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
8700 | - |
nitrocefin | pH 7.4, 22°C, wild-type enzyme | plasmid pBC | |
10000 | - |
nitrocefin | pH 7.4, 22°C, T237S mutant enzyme | plasmid pBC | |
13000 | - |
nitrocefin | pH 7.4, 22°C, T237A mutant enzyme | plasmid pBC |