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Literature summary extracted from

  • Leonard, D.A.; Hujer, A.M.; Smith, B.A.; Schneider, K.D.; Bethel, C.R.; Hujer, K.M.; Bonomo, R.A.
    The role of OXA-1 beta -lactamase Asp66 in the stabilization of the active-site carbamate group and in substrate turnover (2008), Biochem. J., 410, 455-462.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.2.6 expressed in Escherichia coli DH10B Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
3.5.2.6 D66A site-saturation mutagenesis at position 66, 8fold increase in minimum inhibitory concentration for ceftazidime compared with the wild type (0.25 to 2 microg/ml). Mutation results in only minor changes in steady-state protein levels Escherichia coli
3.5.2.6 D66C site-saturation mutagenesis at position 66. Expression of this variant is diminished relative to that of the other proteins Escherichia coli
3.5.2.6 D66L site-saturation mutagenesis at position 66, rise in resistance against ceftazidime compared with the wild type Escherichia coli
3.5.2.6 D66M site-saturation mutagenesis at position 66, rise in resistance against ceftazidime compared with the wild type. Mutation results in only minor changes in steady-state protein levels Escherichia coli
3.5.2.6 D66N site-saturation mutagenesis at position 66, mutation leads to a moderate enhancement of extended-spectrum cephalosporin resistance and an increase in cefepime and cefotaxime resistance. Mutation results in only minor changes in steady-state protein levels Escherichia coli
3.5.2.6 D66P site-saturation mutagenesis at position 66 Escherichia coli
3.5.2.6 D66W site-saturation mutagenesis at position 66, mutation results in only minor changes in steady-state protein levels Escherichia coli
3.5.2.6 D66Y site-saturation mutagenesis at position 66, mutation results in only minor changes in steady-state protein levels Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.2.6 ampicillin
-
 Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.2.6 0.002
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid D66N mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.0024
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid wild type, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.0027
-
 ampicillin D66Y mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.0032
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid D66M mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.006
-
 ampicillin D66A mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.006
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid D66W mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.007
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid D66Y mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.009
-
 cephalothin wild type, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.01
-
 cephalothin D66M mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.011
-
 cephalothin D66N mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.012
-
 ampicillin D66M mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.013
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid D66A mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.014
-
 cefotaxime wild type OXA-1, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.021
-
 ampicillin D66W mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.021
-
 cefotaxime His6-tagged wild type OXA-1, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.021
-
 ampicillin wild type, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.023
-
 ampicillin D66N mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.024
-
 cefotaxime D66N mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.027
-
 cefepime D66A mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.029
-
 ampicillin wild type OXA-1, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.035
-
 cefepime D66M mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.036
-
 cefotaxime D66M mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.036
-
 ampicillin His6-tagged wild type OXA-1, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.036
-
 cefotaxime wild type, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.037
-
 cephalothin D66Y mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.057
-
 cefotaxime D66Y mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.059
-
 cephalothin D66W mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.07
-
 cefotaxime D66A mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.1
-
 cephalothin D66A mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.15
-
 cefepime D66N mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.17
-
 cefepime wild type, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.21
-
 cefepime D66W mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.23
-
 cefepime D66Y mutant, pH 7.4, room temperature Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
3.5.2.6 Escherichia coli P13661
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.2.6 wild type OXA-1 purified by CM-cellulose chromatography and His6-tagged wild type OXA-1 purified by nickel-affinity chromatography Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.2.6 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid + H2O
-
 Escherichia coli ?
-
 ?
3.5.2.6 ampicillin + H2O
-
 Escherichia coli (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 cefepime + H2O
-
 Escherichia coli (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
-
 Escherichia coli (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 cephalothin + H2O
-
 Escherichia coli (2R)-5-[(acetyloxy)methyl]-2-[(R)-carboxy[(thiophen-2-ylacetyl)amino]methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.5.2.6 beta-lactamase
-
 Escherichia coli
3.5.2.6 OXA-1
-
 Escherichia coli
3.5.2.6 oxa-1 beta-lactamase
-
 Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.2.6 0.008
-
 cefepime D66W mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.08
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid D66W mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.15
-
 cephalothin D66W mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.17
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid D66M mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.19
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid D66A mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.25
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid D66Y mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.27
-
 cephalothin D66A mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.28
-
 cephalothin D66Y mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.31
-
 cefotaxime D66M mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.37
-
 cefotaxime D66A mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.38
-
 cephalothin D66M mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.43
-
 cefepime D66M mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.55
-
 cefepime D66A mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 0.71
-
 cefotaxime D66Y mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 1.2
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid D66N mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 1.6
-
 cephalothin D66N mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 1.6
-
 (6R,7S)-3-[(3-carboxy-4-nitro-phenyl)sulfanylmethyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]-oct-2-ene-2-carboxylic acid wild type, pH 7.4, room temperature Escherichia coli
3.5.2.6 2.7
-
 cephalothin wild type, pH 7.4, room temperature Escherichia coli
3.5.2.6 3.5
-
 cefotaxime D66N mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 3.8
-
 cefepime D66Y mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 5.3
-
 ampicillin His6-tagged wild type OXA-1, pH 7.4, room temperature Escherichia coli
3.5.2.6 5.3
-
 cefotaxime wild type, pH 7.4, room temperature Escherichia coli
3.5.2.6 6.7
-
 ampicillin wild type OXA-1, pH 7.4, room temperature Escherichia coli
3.5.2.6 10.6
-
 ampicillin D66Y mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 13
-
 ampicillin D66A mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 27
-
 cefepime D66N mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 34
-
 cefepime wild type, pH 7.4, room temperature Escherichia coli
3.5.2.6 43
-
 ampicillin D66M mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 56
-
 ampicillin D66W mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 440
-
 cefotaxime wild type OXA-1, pH 7.4, room temperature Escherichia coli
3.5.2.6 480
-
 ampicillin D66N mutant, pH 7.4, room temperature Escherichia coli
3.5.2.6 520
-
 cefotaxime His6-tagged wild type OXA-1, pH 7.4, room temperature Escherichia coli
3.5.2.6 520
-
 ampicillin wild type, pH 7.4, room temperature Escherichia coli