Cloned (Comment) | Organism |
---|---|
gene blaSHV-14, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strains BL21(DE3) and CS109. On expression, the clone of wild-type SHV-14 aggravates the resistance of host by 60-500 folds against penicillin and cephalosporin groups of antibiotics, while the expression of mutated enzymes (especially E164A) substantially reduces the resistance level as compared to the wild-type, and the results are in synchrony with the estimated enzymatic efficiencies of wild-type and mutated proteins | Klebsiella pneumoniae |
Protein Variants | Comment | Organism |
---|---|---|
E162A | site-directed mutagenesis, mutation of the an omega loop glutamate, the mutant shows reduced activity against penicilin and cepahlosporin antibiotics in recombinant Escherichia coli cells and reduces the resistance level of the cells | Klebsiella pneumoniae |
E164A | site-directed mutagenesis, mutation of the an omega loop glutamate, the mutant shows highly reduced activity against penicilin and cepahlosporin antibiotics in recombinant Escherichia coli cells and potently reduces the resistance level of the cells | Klebsiella pneumoniae |
E167A | site-directed mutagenesis, mutation of the an omega loop glutamate, the mutant shows reduced activity against penicilin and cepahlosporin antibiotics in recombinant Escherichia coli cells and reduces the resistance level of the cells | Klebsiella pneumoniae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
clavulanic acid | - |
Klebsiella pneumoniae | |
additional information | mutation at the glutamate residues in the omega-loop of SHV-14, i.e. E126, E164, and E167, render beta-lactamase inhibitors more efficient | Klebsiella pneumoniae | |
Sulbactam | - |
Klebsiella pneumoniae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten steady-state kinetics studies | Klebsiella pneumoniae | |
0.032 | - |
ampicillin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
0.038 | - |
nitrocefin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
0.0383 | - |
cefoperazone | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
0.041 | - |
amoxicillin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
0.045 | - |
nitrocefin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
0.045 | - |
cephalothin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
0.05414 | - |
amoxicillin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
0.0557 | - |
piperacillin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
0.057 | - |
cephalothin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
0.0574 | - |
cefoperazone | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
0.058 | - |
piperacillin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
0.0697 | - |
amoxicillin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
0.0716 | - |
piperacillin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
0.0962 | - |
cefoperazone | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
0.1893 | - |
nitrocefin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
0.227 | - |
cephalothin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
0.414 | - |
ampicillin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
0.615 | - |
ampicillin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
amoxicillin + H2O | Klebsiella pneumoniae | - |
(2R,4S)-2-[(R)-[[(2R)-2-amino-2-(4-hydroxyphenyl)acetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
amoxicillin + H2O | Klebsiella pneumoniae MTCC 3384 | - |
(2R,4S)-2-[(R)-[[(2R)-2-amino-2-(4-hydroxyphenyl)acetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
ampicillin + H2O | Klebsiella pneumoniae | - |
(2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
ampicillin + H2O | Klebsiella pneumoniae MTCC 3384 | - |
(2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
cefoperazone + H2O | Klebsiella pneumoniae | - |
(2R)-2-[(R)-carboxy([(2S)-2-[(4-ethyl-2,3-dioxopiperazine-1-carbonyl)amino]-2-(4-hydroxyphenyl)acetyl]amino)methyl]-5-([(1-methyl-1H-tetrazol-5-yl)sulfanyl]methyl)-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
cefotaxime + H2O | Klebsiella pneumoniae | - |
(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 | - |
? | |
cefotaxime + H2O | Klebsiella pneumoniae MTCC 3384 | - |
(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 | - |
? | |
ceftazidime + H2O | Klebsiella pneumoniae | - |
? | - |
? | |
cephalothin + H2O | Klebsiella pneumoniae | - |
(2R)-5-[(acetyloxy)methyl]-2-[(R)-carboxy[2-(thiophen-2-yl)acetamido]methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
imipenem + H2O | Klebsiella pneumoniae | - |
(5R)-3-[[2-(carbonoimidoylamino)ethyl]sulfanyl]-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-4,5-dihydro-1H-pyrrole-2-carboxylic acid | - |
? | |
meropenem + H2O | Klebsiella pneumoniae | - |
(4R,5S)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-3-[[(3S,5S)-5-(dimethylcarbamoyl)pyrrolidin-3-yl]sulfanyl]-4-methyl-4,5-dihydro-1H-pyrrole-2-carboxylic acid | - |
? | |
additional information | Klebsiella pneumoniae | antibiotic sensitivity of recombinant wild-type and mutant blaSHV-14, overview | ? | - |
- |
|
additional information | Klebsiella pneumoniae MTCC 3384 | antibiotic sensitivity of recombinant wild-type and mutant blaSHV-14, overview | ? | - |
- |
|
nitrocefin + H2O | Klebsiella pneumoniae | - |
(2R)-2-((R)-carboxy[2-(thiophen-2-yl)acetamido]methyl)-5-[(E)-2-(2,4-dinitrophenyl)ethenyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
penicillin G + H2O | Klebsiella pneumoniae | - |
(2R,4S)-2-[(R)-carboxy[(phenylacetyl)amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
penicillin G + H2O | Klebsiella pneumoniae MTCC 3384 | - |
(2R,4S)-2-[(R)-carboxy[(phenylacetyl)amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
piperacillin + H2O | Klebsiella pneumoniae | - |
(2R,4S)-2-[(R)-carboxy([(2R)-2-[(4-ethyl-2,3-dioxopiperazine-1-carbonyl)amino]-2-phenylacetyl]amino)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Klebsiella pneumoniae | Q9F918 | - |
- |
Klebsiella pneumoniae MTCC 3384 | Q9F918 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography | Klebsiella pneumoniae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
amoxicillin + H2O | - |
Klebsiella pneumoniae | (2R,4S)-2-[(R)-[[(2R)-2-amino-2-(4-hydroxyphenyl)acetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
amoxicillin + H2O | - |
Klebsiella pneumoniae MTCC 3384 | (2R,4S)-2-[(R)-[[(2R)-2-amino-2-(4-hydroxyphenyl)acetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
ampicilin + H2O | - |
Klebsiella pneumoniae | (2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
ampicillin + H2O | - |
Klebsiella pneumoniae | (2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
ampicillin + H2O | - |
Klebsiella pneumoniae MTCC 3384 | (2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
cefoperazone + H2O | - |
Klebsiella pneumoniae | (2R)-2-[(R)-carboxy([(2S)-2-[(4-ethyl-2,3-dioxopiperazine-1-carbonyl)amino]-2-(4-hydroxyphenyl)acetyl]amino)methyl]-5-([(1-methyl-1H-tetrazol-5-yl)sulfanyl]methyl)-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
cefotaxime + H2O | - |
Klebsiella pneumoniae | (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 | - |
? | |
cefotaxime + H2O | - |
Klebsiella pneumoniae MTCC 3384 | (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 | - |
? | |
ceftazidime + H2O | - |
Klebsiella pneumoniae | ? | - |
? | |
cephalothin + H2O | - |
Klebsiella pneumoniae | (2R)-5-[(acetyloxy)methyl]-2-[(R)-carboxy[2-(thiophen-2-yl)acetamido]methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
imipenem + H2O | - |
Klebsiella pneumoniae | (5R)-3-[[2-(carbonoimidoylamino)ethyl]sulfanyl]-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-4,5-dihydro-1H-pyrrole-2-carboxylic acid | - |
? | |
meropenem + H2O | - |
Klebsiella pneumoniae | (4R,5S)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-3-[[(3S,5S)-5-(dimethylcarbamoyl)pyrrolidin-3-yl]sulfanyl]-4-methyl-4,5-dihydro-1H-pyrrole-2-carboxylic acid | - |
? | |
additional information | antibiotic sensitivity of recombinant wild-type and mutant blaSHV-14, overview | Klebsiella pneumoniae | ? | - |
- |
|
additional information | antibiotic sensitivity of recombinant wild-type and mutant blaSHV-14, overview | Klebsiella pneumoniae MTCC 3384 | ? | - |
- |
|
nitrocefin + H2O | - |
Klebsiella pneumoniae | (2R)-2-((R)-carboxy[2-(thiophen-2-yl)acetamido]methyl)-5-[(E)-2-(2,4-dinitrophenyl)ethenyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
penicillin G + H2O | - |
Klebsiella pneumoniae | (2R,4S)-2-[(R)-carboxy[(phenylacetyl)amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
penicillin G + H2O | - |
Klebsiella pneumoniae MTCC 3384 | (2R,4S)-2-[(R)-carboxy[(phenylacetyl)amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
piperacillin + H2O | - |
Klebsiella pneumoniae | (2R,4S)-2-[(R)-carboxy([(2R)-2-[(4-ethyl-2,3-dioxopiperazine-1-carbonyl)amino]-2-phenylacetyl]amino)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 30000, about, recombinant wild-type and mutant enzymes, SDS-PAGE and MALDI-ToF MS analysis | Klebsiella pneumoniae |
Synonyms | Comment | Organism |
---|---|---|
blaSHV-14 | - |
Klebsiella pneumoniae |
SHV-14 beta-lactamase | - |
Klebsiella pneumoniae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Klebsiella pneumoniae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.08 | - |
nitrocefin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
0.125 | - |
nitrocefin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
0.21 | - |
cefoperazone | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
0.43 | - |
cephalothin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
0.44 | - |
cephalothin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
0.705 | - |
cefoperazone | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
2.03 | - |
amoxicillin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
2.2 | - |
piperacillin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
2.38 | - |
amoxicillin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
2.46 | - |
piperacillin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
5.85 | - |
nitrocefin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
15 | - |
ampicillin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
21.5 | - |
ampicillin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
113 | - |
cephalothin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
229.5 | - |
cefoperazone | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
532.5 | - |
amoxicillin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
544 | - |
ampicillin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
677 | - |
piperacillin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Klebsiella pneumoniae |
General Information | Comment | Organism |
---|---|---|
malfunction | mutation at the glutamate residues in the omega-loop of SHV-14, i.e. E126, E164, and E167, can considerably modulate the beta-lactam sensitivity and reduce hydrolysis. The mutations in SHV-14 render beta-lactamase inhibitors more efficient. The mutations alter the active site configuration of SHV-14 | Klebsiella pneumoniae |
additional information | substrate binding to active site residues of SHV-14 wild-type and mutant enzyme proteins, structures, overview | Klebsiella pneumoniae |
physiological function | extensive production of SHV-14 beta-lactamase makes Klebsiella pneumoniae resistant to beta-lactams. The presence of an omega-loop has been reported to influence the beta-lactamase activity, which is also present in SHV-14 | Klebsiella pneumoniae |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.21 | - |
nitrocefin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
0.66 | - |
nitrocefin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
1.89 | - |
cephalothin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
5.48 | - |
cefoperazone | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
7.33 | - |
cefoperazone | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
7.72 | - |
cephalothin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
30.73 | - |
piperacillin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
34.96 | - |
ampicillin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
36.23 | - |
ampicillin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
43.96 | - |
amoxicillin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
44.17 | - |
piperacillin | pH 7.5, 25°C, recombinant His-tagged mutant E162A | Klebsiella pneumoniae | |
49.51 | - |
amoxicillin | pH 7.5, 25°C, recombinant His-tagged mutant E164A | Klebsiella pneumoniae | |
130 | - |
nitrocefin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
2511.1 | - |
cephalothin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
3998.3 | - |
cefoperazone | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
11672.4 | - |
piperacillin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
17000 | - |
ampicillin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae | |
36334.3 | - |
amoxicillin | pH 7.5, 25°C, recombinant His-tagged wild-type enzyme | Klebsiella pneumoniae |