Protein Variants | Comment | Organism |
---|---|---|
additional information | mutagenesis is performed to better understand the basis of the inhibitor-resistant phenotype in SHV, examination of the role of a second-shell residue, Asn276 | synthetic construct |
N276D | reduced susceptibility to ampicillin/clavulanate while maintaining high-level resistance to ampicillin. Slightly diminished kcat/Km for all substrates tested. 5-fold increase in Ki for clavulanate and a 40% reduction in kinact/KI | synthetic construct |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(6Z)-7-oxo-6-(4H-pyrazolo[1,5-c][1,3]thiazol-2-ylmethylidene)-4-thia-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid | - |
synthetic construct | |
clavulanate | - |
synthetic construct | |
meropenem | - |
synthetic construct |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.021 | - |
nitrocefin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
0.028 | - |
nitrocefin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
0.029 | - |
cephalothin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
0.077 | - |
piperacillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
0.083 | - |
cephalothin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
0.105 | - |
piperacillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
0.134 | - |
ampicillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
0.183 | - |
ampicillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
synthetic construct | - |
using the template blaSHV-1 gene in the phagemid vector pBC SK(-), degeneration of oligonucleotides at Ambler position 276 to make the full complement of amino acid substitutions. After PCR mutagenesis, electroporation of the resultant plasmids into Escherichia coli | - |
Purification (Comment) | Organism |
---|---|
- |
synthetic construct |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ampicillin + H2O | - |
synthetic construct | (2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
cephalothin + H2O | - |
synthetic construct | (2R)-5-[(acetyloxy)methyl]-2-[(R)-carboxy[(thiophen-2-ylacetyl)amino]methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
nitrocefin + H2O | - |
synthetic construct | (2R)-2-[(R)-carboxy[(thiophen-2-ylacetyl)amino]methyl]-5-[(E)-2-(2,4-dinitrophenyl)ethenyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid | - |
? | |
piperacillin + H2O | - |
synthetic construct | (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 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
blaSHV-1 | - |
synthetic construct |
SHV-1 | - |
synthetic construct |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
10 | - |
cephalothin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
14 | - |
cephalothin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
137 | - |
nitrocefin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
237 | - |
nitrocefin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
898 | - |
piperacillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
976 | - |
piperacillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
989 | - |
ampicillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
3100 | - |
ampicillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
120 | - |
cephalothin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
470 | - |
cephalothin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
5000 | - |
nitrocefin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
7000 | - |
ampicillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
9000 | - |
piperacillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
11000 | - |
nitrocefin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
12000 | - |
piperacillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct | |
17000 | - |
ampicillin | pH 7.4, 22°C, wild-type enzyme SHV-1 | synthetic construct |