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Literature summary for 3.5.2.6 extracted from
- Characterisation of the metallo-beta-lactamase VIM-6 and its genetic support (2008), Int. J. Antimicrob. Agents, 32, 446-449.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| construction of the expression system for overproduction of VIM-6 in Escherichia coli | Pseudomonas putida |
| expressed in Escherichia coli | Pseudomonas putida |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| aztreonam | - |
Pseudomonas putida |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
cefepime | above 0.2 | Pseudomonas putida | |
| additional information | - |
ertapenem | above 0.6 | Pseudomonas putida | |
| 0.008 | - |
meropenem | - |
Pseudomonas putida | |
| 0.008 | - |
cefalothin | - |
Pseudomonas putida | |
| 0.01 | - |
cefotaxime | - |
Pseudomonas putida | |
| 0.0125 | - |
Imipenem | - |
Pseudomonas putida | |
| 0.013 | - |
piperacillin | - |
Pseudomonas putida | |
| 0.015 | - |
nitrocefin | - |
Pseudomonas putida | |
| 0.023 | - |
ampicillin | - |
Pseudomonas putida | |
| 0.023 | - |
cefoxitin | - |
Pseudomonas putida | |
| 0.058 | - |
ceftazidime | - |
Pseudomonas putida | |
| 0.2 | - |
cefepime | more than | Pseudomonas putida | |
| 0.6 | - |
ertapenem | more than | Pseudomonas putida | |
| 1.8 | - |
tazobactam | - |
Pseudomonas putida | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Zn2+ | included in assay medium | Pseudomonas putida | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 28370 | - |
mass spectrometry | Pseudomonas putida |
| 28370 | - |
determined by mass spectrometry using a MALDI-TOF-MS instrument | Pseudomonas putida |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| a beta-lactam + H2O | Pseudomonas putida | - |
a substituted beta-amino acid | - |
? | |
| additional information | Pseudomonas putida | enzyme hydrolyses all tested penicillins, cephalosporins and carbapenems with the exception of aztreonam | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas putida | Q8GCU7 | - |
- |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| proteolytic modification | mature enzyme is generated by cleavage of a 26-amino acid signal peptide | Pseudomonas putida |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Pseudomonas putida |
| VIM-6 enzyme is purified from the culture supernatant of 800 ml of stationary-phase culture grown with anion exchange chromatography and gel filtration. | Pseudomonas putida |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| cell culture | strain DU25151/00 | Pseudomonas putida | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.0462 | - |
- |
Pseudomonas putida |
| 46200 | - |
after a 6.3fold purification, the yield is 16.8% and the preparation have a purity above 99% | Pseudomonas putida |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| a beta-lactam + H2O | - |
Pseudomonas putida | a substituted beta-amino acid | - |
? | |
| a penicillin + H2O | - |
Pseudomonas putida | a penicilloic acid | - |
? | |
| ampicillin + H2O | - |
Pseudomonas putida | (2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
| aztreonam + H2O | - |
Pseudomonas putida | [(1S,2S)-1-[[(2Z)-2-(2-ammonio-1,3-thiazol-4-yl)-2-[[(2-carboxypropan-2-yl)oxy]imino]acetyl]amino]-1-carboxypropan-2-yl]sulfamate | - |
? | |
| carbapenem + H2O | - |
Pseudomonas putida | [(2R)-2,3-dihydro-1H-pyrrol-2-yl]acetic acid | - |
? | |
| cefalotin + H2O | - |
Pseudomonas putida | (2R)-5-[(acetyloxy)methyl]-2-{(R)-carboxy[2-(thiophen-2-yl)acetamido]methyl}-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
| cefepime + H2O | - |
Pseudomonas putida | (2R)-2-[(R)-[[(2Z)-2-(2-amino-1,3-thiazol-4-yl)-2-(methoxyimino)acetyl]amino](carboxy)methyl]-5-[(1-methylpyrrolidinium-1-yl)methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylate | - |
? | |
| cefotaxime + H2O | - |
Pseudomonas putida | (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 | - |
? | |
| cefoxitin + H2O | - |
Pseudomonas putida | (2R)-5-[(carbamoyloxy)methyl]-2-[(S)-carboxy(methoxy)[(thiophen-2-ylacetyl)amino]methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
| ceftazidime + H2O | - |
Pseudomonas putida | (2R)-2-[(R)-{[(2Z)-2-(2-amino-1,3-thiazol-4-yl)-2-{[(2-carboxypropan-2-yl)oxy]imino}acetyl]amino}(carboxy)methyl]-5-[(pyridin-1-ium-1-yl)methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylate | - |
? | |
| cephalosporin + H2O | - |
Pseudomonas putida | ? | - |
? | |
| ertapenem + H2O | - |
Pseudomonas putida | (4R,5S)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-3-({(3S,5S)-5-[(3-carboxyphenyl)carbamoyl]pyrrolidin-3-yl}sulfanyl)-4-methyl-4,5-dihydro-1H-pyrrole-2-carboxylic acid | - |
? | |
| imipenem + H2O | - |
Pseudomonas putida | (5R)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-3-({2-[(iminomethyl)amino]ethyl}sulfanyl)-4,5-dihydro-1H-pyrrole-2-carboxylic acid | - |
? | |
| meropenem + H2O | - |
Pseudomonas putida | (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 | enzyme hydrolyses all tested penicillins, cephalosporins and carbapenems with the exception of aztreonam | Pseudomonas putida | ? | - |
? | |
| nitrocefin + H2O | - |
Pseudomonas putida | (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 | - |
? | |
| piperacillin + H2O | - |
Pseudomonas putida | (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 | - |
? | |
| tazobactam + H2O | - |
Pseudomonas putida | (2R,4S,5S)-2-(carboxymethyl)-5-methyl-1,1-dioxo-5-[(1H-1,2,3-triazol-1-yl)methyl]-1lambda6,3-thiazolidine-4-carboxylic acid | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| carbapenemase | - |
Pseudomonas putida |
| metallo-beta-lactamase | - |
Pseudomonas putida |
| metallo-beta-lactamase VIM-6 | - |
Pseudomonas putida |
| VIM-6 | - |
Pseudomonas putida |
| VIM-6 enzyme | - |
Pseudomonas putida |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Pseudomonas putida |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
ertapenem | above 25, enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
| additional information | - |
cefepime | above 8, enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
| additional information | - |
aztreonam | unable to determine as Km is too high | Pseudomonas putida | |
| 7 | - |
cefoxitin | - |
Pseudomonas putida | |
| 7 | - |
cefoxitin | enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
| 8 | - |
cefepime | more than | Pseudomonas putida | |
| 8.2 | - |
tazobactam | - |
Pseudomonas putida | |
| 8.2 | - |
tazobactam | enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
| 12 | - |
ceftazidime | - |
Pseudomonas putida | |
| 12 | - |
ceftazidime | enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
| 25 | - |
ertapenem | more than | Pseudomonas putida | |
| 40 | - |
meropenem | - |
Pseudomonas putida | |
| 40 | - |
meropenem | enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
| 214 | - |
Imipenem | - |
Pseudomonas putida | |
| 214 | - |
Imipenem | enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
| 328 | - |
piperacillin | - |
Pseudomonas putida | |
| 328 | - |
piperacillin | enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
| 330 | - |
nitrocefin | - |
Pseudomonas putida | |
| 330 | - |
nitrocefin | enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
| 398 | - |
cefotaxime | - |
Pseudomonas putida | |
| 398 | - |
cefotaxime | enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
| 440 | - |
cefalothin | - |
Pseudomonas putida | |
| 440 | - |
cefalothin | enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
| 560 | - |
ampicillin | - |
Pseudomonas putida | |
| 560 | - |
ampicillin | enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin | Pseudomonas putida | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 3.6 | - |
aztreonam | inhibitor binding constant (Ki) using 10 microM nitrocefin as indicator substrate | Pseudomonas putida | |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Pseudomonas putida | - |
- |
4.9 |
| Pseudomonas putida | determined by isoelectric focussing | - |
4.9 |
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