Application | Comment | Organism |
---|---|---|
synthesis | production of deuterium- or tritium-labeled substances, mutant enzymes could be applied as NADPH regeneration systems | Pseudomonas stutzeri |
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 |
---|---|---|
A176R | strongly decreased Km for NADP+ | Pseudomonas stutzeri |
E175A | strongly decreased Km for NADP+ | Pseudomonas stutzeri |
E175A/A176R | double mutant, 3.6-fold higher efficiency with NAD+, 1000-fold higher efficiency with NADP+, 3-fold favor for NADP+ over NAD+ as cofactor | Pseudomonas stutzeri |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.004 | - |
NADP+ | 25°C, pH 7.25, E175A/A176R mutant enzyme | Pseudomonas stutzeri | |
0.016 | - |
NAD+ | 25°C, pH 7.25, E175A mutant enzyme | Pseudomonas stutzeri | |
0.02 | - |
NAD+ | 25°C, pH 7.25, E175A/A176R mutant enzyme | Pseudomonas stutzeri | |
0.053 | - |
NAD+ | 25°C, pH 7.25, wild type enzyme | Pseudomonas stutzeri | |
0.06 | - |
NAD+ | 25°C, pH 7.25, A176R mutant enzyme | Pseudomonas stutzeri | |
0.077 | - |
NADP+ | 25°C, pH 7.25, A176R mutant enzyme | Pseudomonas stutzeri | |
0.144 | - |
NADP+ | 25°C, pH 7.25, E175A mutant enzyme | Pseudomonas stutzeri | |
2.51 | - |
NADP+ | 25°C, pH 7.25, wild type enzyme | Pseudomonas stutzeri |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
38500 | - |
x * 38500, SDS-PAGE, His-tag fusion 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 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas stutzeri | - |
- |
- |
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 + NADP+ | 100-fold lower activity than with NAD+ | Pseudomonas stutzeri | phosphate + NADPH | - |
ir |
Subunits | Comment | Organism |
---|---|---|
? | x * 38500, SDS-PAGE, His-tag fusion protein | Pseudomonas stutzeri |
Synonyms | Comment | Organism |
---|---|---|
PTDH | - |
Pseudomonas stutzeri |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40.5 | - |
rapid inactivation, half life less than 10 min, 1 mM NAD+ partially protects against thermal inactivation, E175A/A176R double mutant is readily protected against inactivation by addition of 1 mM NADP+ for at least 15 min | Pseudomonas stutzeri |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.41 | - |
NADP+ | 25°C, pH 7.25, wild type enzyme | Pseudomonas stutzeri | |
1.9 | - |
NADP+ | 25°C, pH 7.25, E175A/A176R mutant enzyme | Pseudomonas stutzeri | |
2.18 | - |
NADP+ | 25°C, pH 7.25, A176R mutant enzyme | Pseudomonas stutzeri | |
2.18 | - |
NADP+ | 25°C, pH 7.25, E175A mutant enzyme | Pseudomonas stutzeri | |
2.93 | - |
NAD+ | 25°C, pH 7.25, wild type enzyme | Pseudomonas stutzeri | |
3.5 | - |
NAD+ | 25°C, pH 7.25, E175A mutant enzyme | Pseudomonas stutzeri | |
3.94 | - |
NAD+ | 25°C, pH 7.25, E175A/A176R mutant enzyme | Pseudomonas stutzeri | |
4.28 | - |
NAD+ | 25°C, pH 7.25, A176R mutant enzyme | Pseudomonas stutzeri |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Pseudomonas stutzeri | |
NADP+ | 100-fold lower activity than with NAD+ | Pseudomonas stutzeri |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Pseudomonas stutzeri | isoelectric focusing, wild type enzyme | - |
5.8 |
Pseudomonas stutzeri | isoelectric focusing, A176R mutant enzyme | - |
6.2 |
Pseudomonas stutzeri | isoelectric focusing, E175A mutant enzyme | - |
6.2 |
Pseudomonas stutzeri | isoelectric focusing, E175A/A176R mutant enzyme | - |
6.6 |