Cloned (Comment) | Organism |
---|---|
PCR-amplification, expression in Escherichia coli DH5alpha, JM109, BL21 (DE3), and Rosetta | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0056 | - |
4-aminoacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.0381 | - |
4-hydroxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.0885 | - |
3-methoxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.0894 | - |
propiophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.103 | - |
2-acetylpyrrole | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.112 | - |
trifluoroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.121 | - |
2-hydroxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.14 | - |
4-chloroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.199 | - |
4-methoxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.208 | - |
4-fluoroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.241 | - |
4-methylacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.244 | - |
3-nitroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.246 | - |
3'-hydroxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.257 | - |
benzaldehyde | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.287 | - |
3-aminoacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.313 | - |
4-Nitroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.393 | - |
acetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.953 | - |
2-decanone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
4.857 | - |
2,4-Pentanedione | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
18.85 | - |
methyl-4-tolyl sulfide | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
21.24 | - |
2-phenylpropionaldehyde | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
30.75 | - |
butyrophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxyacetophenone + NADPH + H+ + O2 | Pseudomonas putida | and related cyclic aliphatic ketones, no activity with NADH as cofactor | 4-hydroxyphenyl acetate + NADP+ + H2O | - |
? | |
4-hydroxyacetophenone + NADPH + H+ + O2 | Pseudomonas putida JD1 | and related cyclic aliphatic ketones, no activity with NADH as cofactor | 4-hydroxyphenyl acetate + NADP+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | B7T912 | JD1 | - |
Pseudomonas putida JD1 | B7T912 | JD1 | - |
Purification (Comment) | Organism |
---|---|
overexpressing Escherichia coli cells are centrifuged, washed, disrupted with a French pressure cell, centrifuged, supernatant (crude extract) purified to homogeneity in 2 chromatographic steps with Q-Sepharose and Reactive Red | Pseudomonas putida |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.54 | - |
crude cell extract, decrease of NADPH, 50 mM Tris-HCl, pH 8.0, 312.5 NADPH, air saturated, 30°C | Pseudomonas putida |
7.5 | - |
Q-Sepharose purified, decrease of NADPH, 50 mM Tris-HCl, pH 8.0, 312.5 NADPH, air saturated, 30°C | Pseudomonas putida |
23.2 | - |
Reactive Red purified, decrease of NADPH, 50 mM Tris-HCl, pH 8.0, 312.5 NADPH, air saturated, 30°C | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2'-hydroxyacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
2'-hydroxyacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida JD1 | ? + NADP+ + H2O | - |
? | |
2,4-pentanedione + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
2-acetylpyrrole + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
2-decanone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
2-hydroxyacetophenone + NADPH + H+ + O2 | - |
Pseudomonas putida | 2-hydroxyphenyl acetate + NADP+ + H2O | - |
? | |
2-hydroxyacetophenone + NADPH + H+ + O2 | - |
Pseudomonas putida JD1 | 2-hydroxyphenyl acetate + NADP+ + H2O | - |
? | |
2-phenylpropionaldehyde + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
3'-hydroxyacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
3-aminoacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
3-methoxyacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
3-nitroacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
3-phenyl-2-butanone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 25°C | Pseudomonas putida | 1-phenylethyl acetate + NADP+ + H2O | - |
? | |
3-phenyl-2-pentanone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 25°C | Pseudomonas putida | 1-phenylpropyl acetate + NADP+ + H2O | - |
? | |
4-aminoacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
4-chloroacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
4-fluoroacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
4-hydroxy-2-decanone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 25°C | Pseudomonas putida | 2-hydroxyoctyl acetate + NADP+ + H2O | - |
? | |
4-hydroxy-2-nonanone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 25°C | Pseudomonas putida | 2-hydroxyheptyl acetate + NADP+ + H2O | - |
? | |
4-hydroxy-2-octanone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 25°C | Pseudomonas putida | 2-hydroxyhexyl acetate + NADP+ + H2O | - |
? | |
4-hydroxy-2-undecanone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 25°C | Pseudomonas putida | 2-hydroxynonyl acetate + NADP+ + H2O | - |
? | |
4-hydroxy-4-phenyl-2-butanone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 25°C | Pseudomonas putida | 1-hydroxy-1-phenylethyl acetate + NADP+ + H2O | - |
? | |
4-hydroxyacetophenone + NADPH + H+ + O2 | and related cyclic aliphatic ketones, no activity with NADH as cofactor | Pseudomonas putida | 4-hydroxyphenyl acetate + NADP+ + H2O | - |
? | |
4-hydroxyacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | 4-hydroxyphenyl acetate + NADP+ + H2O | - |
? | |
4-hydroxyacetophenone + NADPH + H+ + O2 | and related cyclic aliphatic ketones, no activity with NADH as cofactor | Pseudomonas putida JD1 | 4-hydroxyphenyl acetate + NADP+ + H2O | - |
? | |
4-hydroxyacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida JD1 | 4-hydroxyphenyl acetate + NADP+ + H2O | - |
? | |
4-methoxyacetophenone + NADPH + H+ + O2 | - |
Pseudomonas putida | 4-methoxyphenyl acetate + NADP+ + H2O | - |
? | |
4-methoxyacetophenone + NADPH + H+ + O2 | - |
Pseudomonas putida JD1 | 4-methoxyphenyl acetate + NADP+ + H2O | - |
? | |
4-methoxyacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
4-methylacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
4-nitroacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
acetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
benzaldehyde + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
butyrophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
methyl-4-tolyl sulfide + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
propiophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? | |
trifluoroacetophenone + NADPH + H+ + O2 | 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | ? + NADP+ + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
4-hydroxyacetophenone monooxygenase | - |
Pseudomonas putida |
HAPMO | - |
Pseudomonas putida |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 35 | highest thermal stability at 30°C, at 35°C after 30 min 47% activity is retained, at 40°C less than 1%, pH 8.0, 50 mM Tris-HCl buffer | Pseudomonas putida |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.4 | - |
3-methoxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
1 | - |
3-aminoacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
1.2 | - |
4-methoxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
1.2 | - |
3-nitroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
1.3 | - |
4-fluoroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
1.6 | - |
4-chloroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
2.2 | - |
benzaldehyde | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
2.6 | - |
4-Nitroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
2.6 | - |
propiophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
3.5 | - |
2,4-Pentanedione | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
4.2 | - |
2-decanone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
4.4 | - |
4-methylacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
4.9 | - |
butyrophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
8.1 | - |
3'-hydroxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
8.2 | - |
4-aminoacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
8.7 | - |
2-phenylpropionaldehyde | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
9.8 | - |
4-hydroxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
11.3 | - |
2-acetylpyrrole | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
15.4 | - |
trifluoroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
16.3 | - |
acetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
20 | - |
methyl-4-tolyl sulfide | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
21.2 | - |
2'-hydroxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
Tris-HCl, below pH 6.0 the activity is very low | Pseudomonas putida |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 9 | maximum activity pH at 8.0, below pH 6.0 the activity is very low, acetate buffer (pH 4.0-5.5), phosphate buffer (pH 7.5-8.0), Tris-HCl (pH 7.5-9.0), 25°, 30 min | Pseudomonas putida |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Pseudomonas putida |
General Information | Comment | Organism |
---|---|---|
metabolism | recombinant Baeyer-Villiger monooxygenases converts arylaliphatic ketones into the corresponding esters with preference for para-substitution at the aromatic ring, oxidation of aldehydes and some heteroaromatic compounds are also converted while cycloketones and open-chain ketones are not or poorly accepted as substrates, aromatic ketones such as 3-phenyl-2-butanone are oxidized with excellent enantioselectivity | Pseudomonas putida |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.2 | - |
butyrophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.4 | - |
2-phenylpropionaldehyde | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
0.7 | - |
2,4-Pentanedione | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
1.1 | - |
methyl-4-tolyl sulfide | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
3.3 | - |
3-aminoacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
4.3 | - |
3-methoxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
4.4 | - |
2-decanone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
4.9 | - |
3-nitroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
5.9 | - |
4-methoxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
6.2 | - |
4-fluoroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
8.1 | - |
benzaldehyde | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
8.4 | - |
4-Nitroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
11 | - |
4-chloroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
18.5 | - |
4-methylacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
28.5 | - |
propiophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
32.7 | - |
3'-hydroxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
41.6 | - |
acetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
110 | - |
2-acetylpyrrole | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
138 | - |
trifluoroacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
176 | - |
2-hydroxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
257 | - |
4-hydroxyacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida | |
1450 | - |
4-aminoacetophenone | 312 microM NADPH, 50 mM Tris-HCl, pH 8.0, 30°C | Pseudomonas putida |