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Literature summary extracted from

  • Koh, T.H.; Yamaguchi, K.; Ishii, Y.
    Characterisation of the metallo-beta-lactamase VIM-6 and its genetic support (2008), Int. J. Antimicrob. Agents, 32, 446-449.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.2.6 construction of the expression system for overproduction of VIM-6 in Escherichia coli Pseudomonas putida
3.5.2.6 expressed in Escherichia coli Pseudomonas putida

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.2.6 aztreonam
-
 Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.2.6 additional information
-
 cefepime above 0.2 Pseudomonas putida
3.5.2.6 additional information
-
 ertapenem above 0.6 Pseudomonas putida
3.5.2.6 0.008
-
 meropenem
-
 Pseudomonas putida
3.5.2.6 0.008
-
 cefalothin
-
 Pseudomonas putida
3.5.2.6 0.01
-
 cefotaxime
-
 Pseudomonas putida
3.5.2.6 0.0125
-
 Imipenem
-
 Pseudomonas putida
3.5.2.6 0.013
-
 piperacillin
-
 Pseudomonas putida
3.5.2.6 0.015
-
 nitrocefin
-
 Pseudomonas putida
3.5.2.6 0.023
-
 ampicillin
-
 Pseudomonas putida
3.5.2.6 0.023
-
 cefoxitin
-
 Pseudomonas putida
3.5.2.6 0.058
-
 ceftazidime
-
 Pseudomonas putida
3.5.2.6 0.2
-
 cefepime more than Pseudomonas putida
3.5.2.6 0.6
-
 ertapenem more than Pseudomonas putida
3.5.2.6 1.8
-
 tazobactam
-
 Pseudomonas putida

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.2.6 Zn2+ included in assay medium Pseudomonas putida

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.2.6 28370
-
 mass spectrometry Pseudomonas putida
3.5.2.6 28370
-
 determined by mass spectrometry using a MALDI-TOF-MS instrument Pseudomonas putida

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.2.6 a beta-lactam + H2O Pseudomonas putida
-
 a substituted beta-amino acid
-
 ?
3.5.2.6 additional information Pseudomonas putida enzyme hydrolyses all tested penicillins, cephalosporins and carbapenems with the exception of aztreonam ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.2.6 Pseudomonas putida Q8GCU7
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.5.2.6 proteolytic modification mature enzyme is generated by cleavage of a 26-amino acid signal peptide Pseudomonas putida

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.2.6
-
 Pseudomonas putida
3.5.2.6 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

EC Number Source Tissue Comment Organism Textmining
3.5.2.6 cell culture strain DU25151/00 Pseudomonas putida
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.2.6 0.0462
-
-
 Pseudomonas putida
3.5.2.6 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.2.6 a beta-lactam + H2O
-
 Pseudomonas putida a substituted beta-amino acid
-
 ?
3.5.2.6 a penicillin + H2O
-
 Pseudomonas putida a penicilloic acid
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 carbapenem + H2O
-
 Pseudomonas putida [(2R)-2,3-dihydro-1H-pyrrol-2-yl]acetic acid
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 cephalosporin + H2O
-
 Pseudomonas putida ?
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 additional information enzyme hydrolyses all tested penicillins, cephalosporins and carbapenems with the exception of aztreonam Pseudomonas putida ?
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 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

EC Number Synonyms Comment Organism
3.5.2.6 carbapenemase
-
 Pseudomonas putida
3.5.2.6 metallo-beta-lactamase
-
 Pseudomonas putida
3.5.2.6 metallo-beta-lactamase VIM-6
-
 Pseudomonas putida
3.5.2.6 VIM-6
-
 Pseudomonas putida
3.5.2.6 VIM-6 enzyme
-
 Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.2.6 30
-
 assay at Pseudomonas putida

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.2.6 additional information
-
 ertapenem above 25, enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida
3.5.2.6 additional information
-
 cefepime above 8, enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida
3.5.2.6 additional information
-
 aztreonam unable to determine as Km is too high Pseudomonas putida
3.5.2.6 7
-
 cefoxitin
-
 Pseudomonas putida
3.5.2.6 7
-
 cefoxitin enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida
3.5.2.6 8
-
 cefepime more than Pseudomonas putida
3.5.2.6 8.2
-
 tazobactam
-
 Pseudomonas putida
3.5.2.6 8.2
-
 tazobactam enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida
3.5.2.6 12
-
 ceftazidime
-
 Pseudomonas putida
3.5.2.6 12
-
 ceftazidime enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida
3.5.2.6 25
-
 ertapenem more than Pseudomonas putida
3.5.2.6 40
-
 meropenem
-
 Pseudomonas putida
3.5.2.6 40
-
 meropenem enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida
3.5.2.6 214
-
 Imipenem
-
 Pseudomonas putida
3.5.2.6 214
-
 Imipenem enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida
3.5.2.6 328
-
 piperacillin
-
 Pseudomonas putida
3.5.2.6 328
-
 piperacillin enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida
3.5.2.6 330
-
 nitrocefin
-
 Pseudomonas putida
3.5.2.6 330
-
 nitrocefin enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida
3.5.2.6 398
-
 cefotaxime
-
 Pseudomonas putida
3.5.2.6 398
-
 cefotaxime enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida
3.5.2.6 440
-
 cefalothin
-
 Pseudomonas putida
3.5.2.6 440
-
 cefalothin enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida
3.5.2.6 560
-
 ampicillin
-
 Pseudomonas putida
3.5.2.6 560
-
 ampicillin enzyme diluted in HEPES buffer containing 50 microM ZnCl2 supplemented with bovine serum albumin Pseudomonas putida

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.5.2.6 3.6
-
 aztreonam inhibitor binding constant (Ki) using 10 microM nitrocefin as indicator substrate Pseudomonas putida

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.5.2.6 Pseudomonas putida
-
-
 4.9
3.5.2.6 Pseudomonas putida determined by isoelectric focussing
-
 4.9