Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Bacillus cereus |
Crystallization (Comment) | Organism |
---|---|
10 mg/ml purified recombinant wild-type and mutant enzymes, complexed with Mg2+ only or with Mg2+ and inhibitor vinyl sulfonate, in 1 mM HEPES, 10 mM MgCl2, 0.1 mM DTT, pH 7.5, 4°C, hanging drop vapour diffusion method, equal volume of protein and reservoir solution, the latter containing 30% PEG 4000, 100 mM Tris-HCl, pH 7.4, 100 mM MgCl2, 1 week, against the reservoir well solution additionally with 20% glycerol before data collection, X-ray diffraction structure determination and analysis at 2.4-2.8 A resolution | Bacillus cereus |
Protein Variants | Comment | Organism |
---|---|---|
C22A | site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme | Bacillus cereus |
C22S | site-directed mutagenesis, reduced activity compared to the wild-type enzyme | Bacillus cereus |
H56A | site-directed mutagenesis, very highly reduced activity compared to the wild-type enzyme | Bacillus cereus |
M49L | site-directed mutagenesis, very highly reduced activity compared to the wild-type enzyme | Bacillus cereus |
Y128A | site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme | Bacillus cereus |
Y128F | site-directed mutagenesis, reduced activity compared to the wild-type enzyme | Bacillus cereus |
Y128F/C22S | site-directed mutagenesis, reduced activity compared to the wild-type enzyme | Bacillus cereus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
vinyl sulfonate | competitive, inhibition mechanism | Bacillus cereus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.033 | - |
phosphonoacetaldehyde | wild-type enzyme, mutant C22S, and mutant Y128F/C22S, pH 7.5, 25°C | Bacillus cereus | |
0.035 | - |
phosphonoacetaldehyde | mutant Y128A, pH 7.5, 25°C | Bacillus cereus | |
0.045 | - |
phosphonoacetaldehyde | mutant Y128F, pH 7.5, 25°C | Bacillus cereus | |
0.145 | - |
phosphonoacetaldehyde | mutant H56A, pH 7.5, 25°C | Bacillus cereus | |
0.53 | - |
phosphonoacetaldehyde | mutant C22A, pH 7.5, 25°C | Bacillus cereus | |
5.1 | - |
phosphonoacetaldehyde | mutant M49L, pH 7.5, 25°C | Bacillus cereus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required for catalysis | Bacillus cereus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus cereus | O31156 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli | Bacillus cereus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate | mechanism, reaction pathway, Schiff base formation between an amine and a ketone in aqueous solution, active site model | Bacillus cereus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
phosphonoacetaldehyde + H2O | i.e. Pald | Bacillus cereus | acetaldehyde + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
phosphonatase | - |
Bacillus cereus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Bacillus cereus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.075 | - |
phosphonoacetaldehyde | mutant H56A, pH 7.5, 25°C | Bacillus cereus | |
0.077 | - |
phosphonoacetaldehyde | mutant Y128A, pH 7.5, 25°C | Bacillus cereus | |
1.95 | - |
phosphonoacetaldehyde | mutant C22A, pH 7.5, 25°C | Bacillus cereus | |
2.11 | - |
phosphonoacetaldehyde | mutant Y128F/C22S, pH 7.5, 25°C | Bacillus cereus | |
2.21 | - |
phosphonoacetaldehyde | mutant Y128F, pH 7.5, 25°C | Bacillus cereus | |
2.26 | - |
phosphonoacetaldehyde | mutant C22S, pH 7.5, 25°C | Bacillus cereus | |
2.94 | - |
phosphonoacetaldehyde | mutant C22S, pH 7.5, 25°C | Bacillus cereus | |
2.94 | - |
phosphonoacetaldehyde | mutant Y128F, pH 7.5, 25°C | Bacillus cereus | |
2.94 | - |
phosphonoacetaldehyde | mutant Y128F/C22S, pH 7.5, 25°C | Bacillus cereus | |
15 | - |
phosphonoacetaldehyde | wild-type enzyme and mutant M49L, pH 7.5, 25°C | Bacillus cereus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Bacillus cereus |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
1.79 | - |
vinyl sulfonate | pH 7.5, 25°C | Bacillus cereus |