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Literature summary for 3.5.2.6 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
Search for more literature for 3.5.2.6

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli DH10B Escherichia coli

Protein Variants

Protein Variants Comment Organism
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
D66C site-saturation mutagenesis at position 66. Expression of this variant is diminished relative to that of the other proteins Escherichia coli
D66L site-saturation mutagenesis at position 66, rise in resistance against ceftazidime compared with the wild type Escherichia coli
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
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
D66P site-saturation mutagenesis at position 66 Escherichia coli
D66W site-saturation mutagenesis at position 66, mutation results in only minor changes in steady-state protein levels Escherichia coli
D66Y site-saturation mutagenesis at position 66, mutation results in only minor changes in steady-state protein levels Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
ampicillin
-
 Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
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
0.0027
-
 ampicillin D66Y mutant, pH 7.4, room temperature Escherichia coli
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
0.006
-
 ampicillin D66A mutant, pH 7.4, room temperature Escherichia coli
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
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
0.009
-
 cephalothin wild type, pH 7.4, room temperature Escherichia coli
0.01
-
 cephalothin D66M mutant, pH 7.4, room temperature Escherichia coli
0.011
-
 cephalothin D66N mutant, pH 7.4, room temperature Escherichia coli
0.012
-
 ampicillin D66M mutant, pH 7.4, room temperature Escherichia coli
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
0.014
-
 cefotaxime wild type OXA-1, pH 7.4, room temperature Escherichia coli
0.021
-
 ampicillin D66W mutant, pH 7.4, room temperature Escherichia coli
0.021
-
 cefotaxime His6-tagged wild type OXA-1, pH 7.4, room temperature Escherichia coli
0.021
-
 ampicillin wild type, pH 7.4, room temperature Escherichia coli
0.023
-
 ampicillin D66N mutant, pH 7.4, room temperature Escherichia coli
0.024
-
 cefotaxime D66N mutant, pH 7.4, room temperature Escherichia coli
0.027
-
 cefepime D66A mutant, pH 7.4, room temperature Escherichia coli
0.029
-
 ampicillin wild type OXA-1, pH 7.4, room temperature Escherichia coli
0.035
-
 cefepime D66M mutant, pH 7.4, room temperature Escherichia coli
0.036
-
 cefotaxime D66M mutant, pH 7.4, room temperature Escherichia coli
0.036
-
 ampicillin His6-tagged wild type OXA-1, pH 7.4, room temperature Escherichia coli
0.036
-
 cefotaxime wild type, pH 7.4, room temperature Escherichia coli
0.037
-
 cephalothin D66Y mutant, pH 7.4, room temperature Escherichia coli
0.057
-
 cefotaxime D66Y mutant, pH 7.4, room temperature Escherichia coli
0.059
-
 cephalothin D66W mutant, pH 7.4, room temperature Escherichia coli
0.07
-
 cefotaxime D66A mutant, pH 7.4, room temperature Escherichia coli
0.1
-
 cephalothin D66A mutant, pH 7.4, room temperature Escherichia coli
0.15
-
 cefepime D66N mutant, pH 7.4, room temperature Escherichia coli
0.17
-
 cefepime wild type, pH 7.4, room temperature Escherichia coli
0.21
-
 cefepime D66W mutant, pH 7.4, room temperature Escherichia coli
0.23
-
 cefepime D66Y mutant, pH 7.4, room temperature Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P13661
-
-

Purification (Commentary)

Purification (Comment) Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(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 ?
-
 ?
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
-
 ?
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
-
 ?
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
-
 ?
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

Synonyms Comment Organism
beta-lactamase
-
 Escherichia coli
OXA-1
-
 Escherichia coli
oxa-1 beta-lactamase
-
 Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.008
-
 cefepime D66W mutant, pH 7.4, room temperature Escherichia coli
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
0.15
-
 cephalothin D66W mutant, pH 7.4, room temperature Escherichia coli
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
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
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
0.27
-
 cephalothin D66A mutant, pH 7.4, room temperature Escherichia coli
0.28
-
 cephalothin D66Y mutant, pH 7.4, room temperature Escherichia coli
0.31
-
 cefotaxime D66M mutant, pH 7.4, room temperature Escherichia coli
0.37
-
 cefotaxime D66A mutant, pH 7.4, room temperature Escherichia coli
0.38
-
 cephalothin D66M mutant, pH 7.4, room temperature Escherichia coli
0.43
-
 cefepime D66M mutant, pH 7.4, room temperature Escherichia coli
0.55
-
 cefepime D66A mutant, pH 7.4, room temperature Escherichia coli
0.71
-
 cefotaxime D66Y mutant, pH 7.4, room temperature Escherichia coli
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
1.6
-
 cephalothin D66N mutant, pH 7.4, room temperature Escherichia coli
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
2.7
-
 cephalothin wild type, pH 7.4, room temperature Escherichia coli
3.5
-
 cefotaxime D66N mutant, pH 7.4, room temperature Escherichia coli
3.8
-
 cefepime D66Y mutant, pH 7.4, room temperature Escherichia coli
5.3
-
 ampicillin His6-tagged wild type OXA-1, pH 7.4, room temperature Escherichia coli
5.3
-
 cefotaxime wild type, pH 7.4, room temperature Escherichia coli
6.7
-
 ampicillin wild type OXA-1, pH 7.4, room temperature Escherichia coli
10.6
-
 ampicillin D66Y mutant, pH 7.4, room temperature Escherichia coli
13
-
 ampicillin D66A mutant, pH 7.4, room temperature Escherichia coli
27
-
 cefepime D66N mutant, pH 7.4, room temperature Escherichia coli
34
-
 cefepime wild type, pH 7.4, room temperature Escherichia coli
43
-
 ampicillin D66M mutant, pH 7.4, room temperature Escherichia coli
56
-
 ampicillin D66W mutant, pH 7.4, room temperature Escherichia coli
440
-
 cefotaxime wild type OXA-1, pH 7.4, room temperature Escherichia coli
480
-
 ampicillin D66N mutant, pH 7.4, room temperature Escherichia coli
520
-
 cefotaxime His6-tagged wild type OXA-1, pH 7.4, room temperature Escherichia coli
520
-
 ampicillin wild type, pH 7.4, room temperature Escherichia coli