Application | Comment | Organism |
---|---|---|
synthesis | the enzyme is intersting for regioselective production of compounds | Bacillus subtilis |
synthesis | the enzyme is intersting for regioselective production of compounds | Priestia megaterium |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli in fed-batch fermentation | Bacillus subtilis |
expression in Escherichia coli in fed-batch fermentation | Priestia megaterium |
Crystallization (Comment) | Organism |
---|---|
crystallization of the wild-type and mutant CYP102A1 with and without bound substrates and one including theFMNbinding domain | Priestia megaterium |
Protein Variants | Comment | Organism |
---|---|---|
A74E/F87V/P386S | site-directed mutagenesis, the mutant shows altered regioselectivity and activity, and cofactor specificity compared to the wild-type mutant | Priestia megaterium |
A74G/F87V/L188Q | site-directed mutagenesis, the mutant shows altered regioselectivity and activity compared to the wild-type mutant | Priestia megaterium |
A74G/F87V/L188Q/R966D | site-directed mutagenesis, the mutant shows altered kinetics, and cofactor specificity compared to the wild-type enzyme | Priestia megaterium |
A74G/F87V/L188Q/R966M | site-directed mutagenesis, the mutant shows altered kinetics, and cofactor specificity compared to the wild-type enzyme | Priestia megaterium |
A74G/F87V/L188Q/S965D | site-directed mutagenesis, the mutant shows altered kinetics, and cofactor specificity compared to the wild-type enzyme | Priestia megaterium |
A74G/F87V/L188Q/W1046A | site-directed mutagenesis, the mutant shows altered kinetics, and cofactor specificity compared to the wild-type enzyme | Priestia megaterium |
A74G/F87V/L188Q/W1046S | site-directed mutagenesis, the mutant shows altered kinetics, and cofactor specificity compared to the wild-type enzyme | Priestia megaterium |
F87G | site-directed mutagenesis, the mutant shows altered regioselectivity and activity compared to the wild-type mutant | Priestia megaterium |
F87GA | site-directed mutagenesis, the mutant shows altered regioselectivity and activity compared to the wild-type mutant | Priestia megaterium |
additional information | construction of an engineered CYP102A1 heme domain which utilizes H2O2 as electron donor instead of NADPH | Priestia megaterium |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
H2O2 | at high concentrations | Priestia megaterium |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.002 | - |
NADPH | mutants A74G/F87V/L188Q/W1046A and A74G/F87V/L188Q/W1046S | Priestia megaterium | |
0.0025 | - |
NADPH | mutant A74G/F87V/L188Q | Priestia megaterium | |
0.004 | - |
NADH | mutant A74G/F87V/L188Q/S965D | Priestia megaterium | |
0.004 | - |
NADH | mutant enzyme A74G/F87V/L188Q/W1046A/S965D, pH and temperature not specified in the publication | Priestia megaterium | |
0.01 | - |
NADH | mutant A74G/F87V/L188Q/W1046S | Priestia megaterium | |
0.01 | - |
NADH | mutant enzyme A74G/F87V/L188Q/W1046S, pH and temperature not specified in the publication | Priestia megaterium | |
0.02 | - |
NADPH | mutant A74G/F87V/L188Q/S965D | Priestia megaterium | |
0.02 | - |
NADH | mutant A74G/F87V/L188Q/W1046A | Priestia megaterium | |
0.02 | - |
NADH | mutant enzyme A74G/F87V/L188Q/W1046A, pH and temperature not specified in the publication | Priestia megaterium | |
0.05 | - |
NADPH | mutant A74G/F87V/L188Q/R966M | Priestia megaterium | |
0.13 | - |
NADPH | mutant A74G/F87V/L188Q/R966D | Priestia megaterium | |
0.33 | - |
NADH | mutant A74G/F87V/L188Q/R966D | Priestia megaterium | |
0.33 | - |
NADH | mutant enzyme A74G/F87V/L188Q/R966D, pH and temperature not specified in the publication | Priestia megaterium | |
0.54 | - |
NADH | mutant A74G/F87V/L188Q/R966M | Priestia megaterium | |
0.54 | - |
NADH | mutant enzyme A74G/F87V/L188Q/R966M, pH and temperature not specified in the publication | Priestia megaterium | |
1.43 | - |
NADH | mutant A74G/F87V/L188Q | Priestia megaterium | |
1.43 | - |
NADH | mutant enzyme A74G/F87V/L188Q, pH and temperature not specified in the publication | Priestia megaterium |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | a heme-containing P450 monooxygenase | Bacillus subtilis | |
Fe2+ | a heme-containing P450 monooxygenase | Priestia megaterium |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | - |
- |
- |
Priestia megaterium | - |
- |
- |
Purification (Comment) | Organism |
---|---|
several purification protocols are published, based on ion exchange chromatography, hydrophobic interaction chromatography or metal affinity chromatography, resulting in high levels of purity depending on the further application of the enzymes | Bacillus subtilis |
several purification protocols are published, based on ion exchange chromatography, hydrophobic interaction chromatography or metal affinity chromatography, resulting in high levels of purity depending on the further application of the enzymes | Priestia megaterium |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
beta-ionone + [reduced NADPH-hemoprotein reductase] + O2 | - |
Priestia megaterium | 4-hydroxy-beta-ionone + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
cytochrome c + NADH + H+ + O2 | - |
Priestia megaterium | ? | - |
r | |
cytochrome c + O2 + NADH | - |
Priestia megaterium | ? | - |
r | |
additional information | activity involves cytochrome c reduction, CYP102A1 hydroxylates and epoxidizes middle to long chain saturated, unsaturated and branched fatty acids at subterminal positions, an engineered CYP102A1 heme domain which utilizes H2O2 as electron donor | Priestia megaterium | ? | - |
? | |
additional information | activity involves cytochrome c reduction, CYP102A3 hydroxylates and epoxidizes middle to long chain saturated, unsaturated and branched fatty acids at subterminal positions | Bacillus subtilis | ? | - |
? | |
styrene + O2 + H2O2 | engineered CYP102A1 heme domain which utilizes H2O2 as electron donor instead of NADPH | Priestia megaterium | (R)-styrene oxide + (S)-styrene oxide + H2O | - |
? | |
styrene + O2 + NAD(P)H | - |
Priestia megaterium | (R)-styrene oxide + (S)-styrene oxide + H2O + NAD(P)+ | enantioselective styrene oxidation with different CYP102A1 mutants, 25% S-isomer for the wild-type enzyme, 58% S-isomer for mutant A74E/F87V/P386S, 49% R-isomer for mutant F87A, 65% R-isomer for mutant A74G/F87V/L188Q, and 92% R-isomer for mutant F87G | ? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme is a natural fusion enzyme composed of an N-terminal heme monooxygenase linked to the C-terminal diflavin reductase domain | Bacillus subtilis |
More | the enzyme is a natural fusion enzyme composed of an N-terminal heme monooxygenase linked to the C-terminal diflavin reductase domain | Priestia megaterium |
Synonyms | Comment | Organism |
---|---|---|
CYP102 monooxygenase | - |
Bacillus subtilis |
CYP102 monooxygenase | - |
Priestia megaterium |
CYP102A1 | - |
Priestia megaterium |
CYP102A3 | - |
Bacillus subtilis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
melting temperatures of the CYP102A1 monooxygenase and the CYP102A1 reductase domain differ by about 15°C: whereas Tm for the monooxygenase domain is 63°C, it is only 48°C for the reductase domain | Priestia megaterium |
46 | - |
10 min, half-life, wild-type CYP102A1 | Priestia megaterium |
61 | - |
10 min, half-life, engineered CYP102A1 heme domain which utilizes H2O2 as electron donor | Priestia megaterium |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.016 | - |
NADH | mutant A74G/F87V/L188Q | Priestia megaterium | |
0.43 | - |
NADPH | mutant F87A | Priestia megaterium | |
0.45 | - |
NADPH | wild-type enzyme | Priestia megaterium | |
0.5 | - |
NADPH | mutant F87G | Priestia megaterium | |
0.83 | - |
NADPH | mutant A74E/F87V/P386S | Priestia megaterium | |
1.7 | - |
NADPH | mutant A74G/F87V/L188Q | Priestia megaterium | |
5.67 | - |
NADPH | mutant A74G/F87V/L188Q/S965D | Priestia megaterium | |
6.5 | - |
NADPH | mutant A74G/F87V/L188Q/W1046A | Priestia megaterium | |
7 | - |
NADH | mutant A74G/F87V/L188Q/S965D | Priestia megaterium | |
9.3 | - |
NADPH | mutant A74G/F87V/L188Q/R966M | Priestia megaterium | |
9.83 | - |
NADPH | mutant A74G/F87V/L188Q/W1046S | Priestia megaterium | |
32 | - |
NADH | mutant A74G/F87V/L188Q/R966D | Priestia megaterium | |
37.83 | - |
NADH | mutant A74G/F87V/L188Q/R966M | Priestia megaterium | |
46.83 | - |
NADH | mutant A74G/F87V/L188Q | Priestia megaterium | |
89 | - |
NADH | mutant A74G/F87V/L188Q/W1046A | Priestia megaterium | |
105.2 | - |
NADH | mutant A74G/F87V/L188Q/W1046A | Priestia megaterium | |
132.2 | - |
NADPH | mutant A74G/F87V/L188Q | Priestia megaterium | |
156 | - |
NADH | mutant A74G/F87V/L188Q/W1046S | Priestia megaterium | |
183 | - |
NADPH | mutant A74G/F87V/L188Q/R966D | Priestia megaterium | |
362.8 | - |
NADPH | mutant A74G/F87V/L188Q/R966M | Priestia megaterium |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
cytochrome c | - |
Bacillus subtilis | |
cytochrome c | - |
Priestia megaterium | |
FAD | bound | Bacillus subtilis | |
FAD | bound, binding structure, overview | Priestia megaterium | |
FMN | bound | Bacillus subtilis | |
FMN | bound, binding structure, overview | Priestia megaterium | |
additional information | engineered CYP102A1 heme domain which utilizes H2O2 as electron donor instead of NADPH | Priestia megaterium | |
additional information | residues S965, R966 and Y974 are important in the Rossman fold-like binding motif for the cofactor binding and discrimination, overview | Priestia megaterium | |
NAD(P)H | - |
Priestia megaterium | |
NADPH | - |
Bacillus subtilis |