Cloned (Comment) | Organism |
---|---|
wild type and mutant proteins expressed in Escherichia coli BL21(DE3) as His-tag fusion protein | Pseudomonas stutzeri |
Protein Variants | Comment | Organism |
---|---|---|
E266Q | significant increase in Km for both substrates, increase in turnover | Pseudomonas stutzeri |
H292F | almost complete loss of activity | Pseudomonas stutzeri |
H292K | almost complete loss of activity | Pseudomonas stutzeri |
H292N | almost complete loss of activity | Pseudomonas stutzeri |
K76A | significant increase in Km for both substrates | Pseudomonas stutzeri |
K76C | significant increase in Km for both substrates | Pseudomonas stutzeri |
K76M | significant increase in Km for both substrates | Pseudomonas stutzeri |
K76R | significant increase in Km for both substrates | Pseudomonas stutzeri |
R237H | almost complete loss of activity | Pseudomonas stutzeri |
R237K | significant increase in Km for both substrates | Pseudomonas stutzeri |
R237L | almost complete loss of activity | Pseudomonas stutzeri |
R237Q | almost complete loss of activity | Pseudomonas stutzeri |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-Bromoethylamine | 1 mM NAD+ protects against inactivation | Pseudomonas stutzeri | |
diethyl dicarbonate | - |
Pseudomonas stutzeri |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.053 | - |
phosphonate | pH 7.5, 25°C, wild type enzyme | Pseudomonas stutzeri | |
0.057 | - |
NAD+ | pH 7.5, 25°C, wild type enzyme | Pseudomonas stutzeri | |
0.11 | - |
NAD+ | pH 7.5, 25°C, K76R mutant enzyme | Pseudomonas stutzeri | |
0.2 | - |
NAD+ | pH 7.5, 25°C, K76C mutant enzyme | Pseudomonas stutzeri | |
0.26 | - |
NAD+ | pH 7.5, 25°C, K76A mutant enzyme | Pseudomonas stutzeri | |
0.35 | - |
phosphonate | pH 7.5, 25°C, K76R mutant enzyme | Pseudomonas stutzeri | |
0.6 | - |
phosphonate | pH 7.5, 25°C, K76C mutant enzyme | Pseudomonas stutzeri | |
0.9 | - |
NAD+ | pH 7.5, 25°C, K76M mutant enzyme | Pseudomonas stutzeri | |
1 | - |
NAD+ | pH 7.5, 25°C, E266Q mutant enzyme | Pseudomonas stutzeri | |
1 | - |
NAD+ | pH 7.5, 25°C, R237K mutant enzyme | Pseudomonas stutzeri | |
1.1 | - |
phosphonate | pH 7.5, 25°C, K76A mutant enzyme | Pseudomonas stutzeri | |
2 | - |
phosphonate | pH 7.5, 25°C, K76M mutant enzyme | Pseudomonas stutzeri | |
5 | - |
phosphonate | pH 7.5, 25°C, E266Q mutant enzyme | Pseudomonas stutzeri | |
6 | - |
phosphonate | pH 7.5, 25°C, R237K mutant enzyme | Pseudomonas stutzeri |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
36400 | - |
x * 38500, SDS-PAGE, His-tag fusion protein, x * 36400, SDS-PAGE, native protein | Pseudomonas stutzeri |
38500 | - |
x * 38500, SDS-PAGE, His-tag fusion protein, x * 36400, SDS-PAGE, native protein | Pseudomonas stutzeri |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
phosphonate + H2O + NAD+ | Pseudomonas stutzeri | NADH regeneration system | phosphate + NADH | - |
ir | |
phosphonate + H2O + NAD+ | Pseudomonas stutzeri WM 88 | NADH regeneration system | phosphate + NADH | - |
ir |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas stutzeri | - |
- |
- |
Pseudomonas stutzeri WM 88 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzymes using His-tag | Pseudomonas stutzeri |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
phosphonate + H2O + NAD+ | - |
Pseudomonas stutzeri | phosphate + NADH | - |
ir | |
phosphonate + H2O + NAD+ | NADH regeneration system | Pseudomonas stutzeri | phosphate + NADH | - |
ir | |
phosphonate + H2O + NAD+ | - |
Pseudomonas stutzeri WM 88 | phosphate + NADH | - |
ir | |
phosphonate + H2O + NAD+ | NADH regeneration system | Pseudomonas stutzeri WM 88 | phosphate + NADH | - |
ir |
Subunits | Comment | Organism |
---|---|---|
? | x * 38500, SDS-PAGE, His-tag fusion protein, x * 36400, SDS-PAGE, native protein | Pseudomonas stutzeri |
Synonyms | Comment | Organism |
---|---|---|
PTDH | - |
Pseudomonas stutzeri |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.28 | - |
phosphonate | pH 7.5, 25°C, R237K mutant enzyme | Pseudomonas stutzeri | |
0.58 | - |
phosphonate | pH 7.5, 25°C, K76C mutant enzyme | Pseudomonas stutzeri | |
0.82 | - |
phosphonate | pH 7.5, 25°C, K76A mutant enzyme | Pseudomonas stutzeri | |
1.62 | - |
phosphonate | pH 7.5, 25°C, K76M mutant enzyme | Pseudomonas stutzeri | |
2.33 | - |
phosphonate | pH 7.5, 25°C, K76R mutant enzyme | Pseudomonas stutzeri | |
3.2 | - |
phosphonate | pH 7.5, 25°C, wild type enzyme | Pseudomonas stutzeri | |
8 | - |
phosphonate | pH 7.5, 25°C, E266Q mutant enzyme | Pseudomonas stutzeri |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Pseudomonas stutzeri |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 9.5 | - |
Pseudomonas stutzeri |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Pseudomonas stutzeri |