Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant R234K in Escherichia coli BL21(DE3) | Pseudomonas aeruginosa |
Crystallization (Comment) | Organism |
---|---|
0.003 ml 10 mg/ml purified recombinant wild-type enzyme or mutant R234K, mixed with 0.001 ml of well solution containing 2 M ammonium sulfate, 50 mM MOPS pH 6.4, and 0.1 M acetate pH 4.5, respectively, 100 mM MgCl2, 1-2 weeks at room temperature, X-ray diffraction structure determination and analysis at 1.75 A resolution | Pseudomonas aeruginosa |
Protein Variants | Comment | Organism |
---|---|---|
R234K | site-directed replacement mutagenesis, about 4fold reduced catalytic efficiency compared to the wild-type enzyme | Pseudomonas aeruginosa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.033 | - |
ampicillin | pH 7.0, 30°C, wild-type enzyme | Pseudomonas aeruginosa | |
0.057 | - |
penicillin G | pH 7.0, 30°C, wild-type enzyme | Pseudomonas aeruginosa | |
0.068 | - |
carbenicillin | pH 7.0, 30°C, wild-type enzyme | Pseudomonas aeruginosa | |
0.074 | - |
ampicillin | pH 7.0, 30°C, mutant R234K | Pseudomonas aeruginosa | |
0.1 | - |
penicillin G | pH 7.0, 30°C, mutant R234K | Pseudomonas aeruginosa | |
0.129 | - |
carbenicillin | pH 7.0, 30°C, mutant R234K | Pseudomonas aeruginosa |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | P16897 | class A enzyme PSE-4, the active site structure involves Ser70, Lys73, Ser130, Glu166, Asn170, and Arg234 as catalytic residues | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant R234K from Escherichia coli BL21(DE3) | Pseudomonas aeruginosa |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
a beta-lactam + H2O = a substituted beta-amino acid | Arg234 is involved in catalysis | Pseudomonas aeruginosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ampicillin + H2O | - |
Pseudomonas aeruginosa | (2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
carbenicillin + H2O | - |
Pseudomonas aeruginosa | (2R,4S)-2-{(R)-carboxy[2-carboxy(phenyl)acetamido]methyl}-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? | |
nitrocefin + H2O | - |
Pseudomonas aeruginosa | (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 | - |
? | |
penicillin G + H2O | - |
Pseudomonas aeruginosa | (2R,4S)-2-[(R)-carboxy[(phenylacetyl)amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | enzyme consists of an alpha/beta domain and a predominantly alpha-helical domain | Pseudomonas aeruginosa |
Synonyms | Comment | Organism |
---|---|---|
Carbenicillinase | - |
Pseudomonas aeruginosa |
PSE-4 | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Pseudomonas aeruginosa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
57 | - |
carbenicillin | pH 7.0, 30°C, mutant R234K | Pseudomonas aeruginosa | |
430 | - |
penicillin G | pH 7.0, 30°C, mutant R234K | Pseudomonas aeruginosa | |
840 | - |
ampicillin | pH 7.0, 30°C, mutant R234K | Pseudomonas aeruginosa | |
890 | - |
penicillin G | pH 7.0, 30°C, wild-type enzyme | Pseudomonas aeruginosa | |
1170 | - |
ampicillin | pH 7.0, 30°C, wild-type enzyme | Pseudomonas aeruginosa | |
1200 | - |
carbenicillin | pH 7.0, 30°C, wild-type enzyme | Pseudomonas aeruginosa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Pseudomonas aeruginosa |