Cloned (Comment) | Organism |
---|---|
hapE gene, expression in Escherichia coli TOP10 | Pseudomonas fluorescens |
Protein Variants | Comment | Organism |
---|---|---|
K439A | mutant with 100fold decrease in catalytic efficiency with NADPH, mainly caused by increased Km, 4fold increased efficiency with NADH | Pseudomonas fluorescens |
K439F | mutant with higher activity with NADH compared to wild-type HAPMO | Pseudomonas fluorescens |
K439N | mutant with higher activity with NADH compared to wild-type HAPMO | Pseudomonas fluorescens |
K439P | mutant with higher activity with NADH compared to wild-type HAPMO | Pseudomonas fluorescens |
R339A | mutant with largely decreased affinity for NADPH and decreased catalytic efficiency with NADH | Pseudomonas fluorescens |
R440A | totally inactive mutant, Arg-440 is crucial for completing the catalytic cycle | Pseudomonas fluorescens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
acetyl-NADP+ | 0.5 mM, 35% inhibition | Pseudomonas fluorescens | |
amino-NADP+ | very effective inhibitor, 0.005 mM, 80% inhibition | Pseudomonas fluorescens | |
additional information | not inhibited by NADP+ | Pseudomonas fluorescens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.012 | - |
NADPH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, wild-type HAPMO | Pseudomonas fluorescens | |
0.017 | - |
NADPH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439P mutant | Pseudomonas fluorescens | |
0.078 | - |
NADPH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439N mutant | Pseudomonas fluorescens | |
0.08 | - |
NADPH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439F mutant | Pseudomonas fluorescens | |
0.2 | - |
NADPH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439A mutant | Pseudomonas fluorescens | |
0.3 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439F mutant | Pseudomonas fluorescens | |
0.52 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439A mutant | Pseudomonas fluorescens | |
0.78 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439N mutant | Pseudomonas fluorescens | |
1.28 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439P mutant | Pseudomonas fluorescens | |
1.44 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, wild-type HAPMO | Pseudomonas fluorescens | |
3 | - |
NADPH | above, pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, R339A mutant | Pseudomonas fluorescens | |
3 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, R339A mutant | Pseudomonas fluorescens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
145000 | - |
- |
Pseudomonas fluorescens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas fluorescens | - |
- |
- |
Pseudomonas fluorescens ACB | - |
- |
- |
Purification (Comment) | Organism |
---|---|
wild-type and mutant HAPMO | Pseudomonas fluorescens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxyacetophenone + NADH + H+ + O2 | 700fold preference for NADPH over NADH | Pseudomonas fluorescens | 4-hydroxyphenyl acetate + NAD+ + H2O | - |
? | |
4-hydroxyacetophenone + NADH + H+ + O2 | 700fold preference for NADPH over NADH | Pseudomonas fluorescens ACB | 4-hydroxyphenyl acetate + NAD+ + H2O | - |
? | |
4-hydroxyacetophenone + NADPH + H+ + O2 | 700fold preference for NADPH over NADH, Arg-440 plays an important role in catalysis | Pseudomonas fluorescens | 4-hydroxyphenyl acetate + NADP+ + H2O | - |
? | |
4-hydroxyacetophenone + NADPH + H+ + O2 | 700fold preference for NADPH over NADH, Arg-440 plays an important role in catalysis | Pseudomonas fluorescens ACB | 4-hydroxyphenyl acetate + NADP+ + H2O | - |
? | |
additional information | catalyzes Baeyer-Villiger oxidation reactions on various ketones, oxidizes a variety of aromatic ketones and sulfides | Pseudomonas fluorescens | ? | - |
? | |
additional information | catalyzes Baeyer-Villiger oxidation reactions on various ketones, oxidizes a variety of aromatic ketones and sulfides | Pseudomonas fluorescens ACB | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | - |
Pseudomonas fluorescens |
Synonyms | Comment | Organism |
---|---|---|
HAPMO | - |
Pseudomonas fluorescens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Pseudomonas fluorescens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
NADPH | above, pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, R339A mutant | Pseudomonas fluorescens | |
0.19 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, R339A mutant | Pseudomonas fluorescens | |
0.2 | - |
NADPH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439P mutant | Pseudomonas fluorescens | |
0.7 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439P mutant | Pseudomonas fluorescens | |
1 | - |
NADPH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439F mutant | Pseudomonas fluorescens | |
1.6 | - |
NADPH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439A mutant | Pseudomonas fluorescens | |
1.7 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, wild-type HAPMO | Pseudomonas fluorescens | |
2.4 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439A mutant | Pseudomonas fluorescens | |
2.4 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439F mutant | Pseudomonas fluorescens | |
3.5 | - |
NADPH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439N mutant | Pseudomonas fluorescens | |
5.1 | - |
NADH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439N mutant | Pseudomonas fluorescens | |
10.1 | - |
NADPH | pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, wild-type HAPMO | Pseudomonas fluorescens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Pseudomonas fluorescens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | contains one FAD molecule per subunit | Pseudomonas fluorescens | |
NADH | 700fold preference for NADPH over NADH | Pseudomonas fluorescens | |
NADPH | specific for NADPH as coenzyme, Arg-339 and Lys-439 are involved in coenzyme recognition, Arg-440 not, Lys-439 plays a role in recognizing the 2-phosphate of NADPH, 700fold preference for NADPH over NADH | Pseudomonas fluorescens |