Cloned (Comment) | Organism |
---|---|
- |
Pseudomonas putida |
Crystallization (Comment) | Organism |
---|---|
crystal structure of the enzyme complexed with a 1-methylguanidine. All subunits in the complex exist as the closed form. X-ray crystallographic study and the atomic absorption spectrometry analysis of the E122Q mutant-substrate complex reveal that the drastic decrease of the activity of the E122Q is caused by the loss of one Zn ion at the Metal1 site and a critical function of Glu122 | Pseudomonas putida |
Protein Variants | Comment | Organism |
---|---|---|
E122Q | almost complete loss of activity | Pseudomonas putida |
E183Q | complete loss of activity | Pseudomonas putida |
H178A | complete loss of activity | Pseudomonas putida |
W154A | complete loss of activity | Pseudomonas putida |
W154F | almost complete loss of activity | Pseudomonas putida |
W174A | almost complete loss of activity | Pseudomonas putida |
W174F | about 50% of wild-type activity | Pseudomonas putida |
Y121A | about 10% of wild-type catalytic efficiency | Pseudomonas putida |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | wild-type enzyme, purified in the presence of EDTA, shows only half the activity of the wild-type enzyme, the activity is recovered by the addition of ZnCl2 or MnCl2 | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
44 | - |
creatinine | wild-type, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
56 | - |
creatinine | mutant E122Q, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
77 | - |
creatinine | wild-type, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
90 | - |
creatinine | mutant W174F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
150 | - |
creatinine | wild-type, presence of EDTA, semi-apo-enzyme, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
160 | - |
creatinine | mutant Y121A, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
220 | - |
creatinine | mutant E122Q, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
350 | - |
creatinine | mutant W154F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zinc | removal of the zinc ions by dialyzing against buffers containing o-phenanthroline results in a partial decrease of 10-30% in activity and zinc content. Wild-type enzyme, purified in the presence of EDTA, shows only half the activity of the wild-type enzyme, the activity is recovered by the addition of ZnCl2 or MnCl2 | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | P83772 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
creatinine + H2O = creatine | catalytic mechanism: a first water molecule is a hydroxide ion that is bound as a bridge between the two metal ions and attacks the carbonyl carbon of the substrate. A second water molecule is bound to the carboxyl group of Glu122 and functions as a proton donor in catalysis | Pseudomonas putida |
Renatured (Comment) | Organism |
---|---|
wild-type enzyme, purified in the presence of EDTA, shows only half the activity of the wild-type enzyme, the activity is recovered by the addition of ZnCl2 or MnCl2 | Pseudomonas putida |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.13 | - |
mutant E122Q, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
4.9 | - |
mutant W154F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
13.9 | - |
mutant E122Q, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
140 | - |
mutant Y121A, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
178 | - |
wild-type, presence of EDTA, semi-apo-enzyme, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
370 | - |
wild-type, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
740 | - |
mutant W174F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
1560 | - |
wild-type, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
creatinine + H2O | - |
Pseudomonas putida | creatine | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.5 | - |
creatinine | mutant E122Q, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
11 | - |
creatinine | mutant E122Q, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
18 | - |
creatinine | mutant W154F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
86 | - |
creatinine | mutant Y121A, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
222 | - |
creatinine | wild-type, presence of EDTA, semi-apo-enzyme, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
252 | - |
creatinine | wild-type, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
688 | - |
creatinine | mutant W174F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
1340 | - |
creatinine | wild-type, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.008 | - |
creatinine | mutant E122Q, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
0.051 | - |
creatinine | mutant W154F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
0.2 | - |
creatinine | mutant E122Q, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
0.54 | - |
creatinine | mutant Y121A, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
1.48 | - |
creatinine | wild-type, presence of EDTA, semi-apo-enzyme, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
5.73 | - |
creatinine | wild-type, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
7.64 | - |
creatinine | mutant W174F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
17.4 | - |
creatinine | wild-type, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |