EC Number | Cloned (Comment) | Organism |
---|---|---|
1.14.99.15 | overexpression of the His-tagged enzyme in Escherichia coli BL21(DE3) | Rhodopseudomonas palustris |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.14.99.15 | purified recombinant CYP199A2, 16 ºC, the hanging drop vapor diffusion method under aerobic conditions, 0.0015 ml of protein solution is mixed with 0.0015 ml of reservoir solution, addition of 200 ml reservoir solution, containing 15% PEG 4000, 100 mM sodium citrate pH 5.6, 20% isopropanol with 4% v/v t-butanol, 1 week, X-ray diffraction structure deternnation and analysis at 2.0 A resolution | Rhodopseudomonas palustris |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.14.99.15 | additional information | - |
additional information | steady-state kinetics | Rhodopseudomonas palustris |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.14.99.15 | Fe2+ | heme iron | Rhodopseudomonas palustris |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.99.15 | 4-methoxybenzoate + reduced ferredoxin + O2 | Rhodopseudomonas palustris | - |
4-hydroxybenzoate + formaldehyde + ferredoxin + H2O | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.14.99.15 | Rhodopseudomonas palustris | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.14.99.15 | recombinant His-tagged enzyme from Escherichia coli BL21(DE3) by nickel affinity chromatography and gel filtration | Rhodopseudomonas palustris |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.99.15 | 4-ethylbenzoate + AH2 + O2 | a C-C bond dehydrogenation of an unbranched alkyl group, computational docking of 4-ethylbenzoate into the active site suggests that the substrate carboxylate oxygens interact with Ser97 and Ser247, and the beta-methyl group is located over the heme iron by Phe185, this binding orientation is consistent with the observed product profile of exclusive attack at the para substituent, overview | Rhodopseudomonas palustris | 4-(1-hydroxyethyl)-benzoate + 4-vinylbenzoate + A + H2O | - |
? | |
1.14.99.15 | 4-methoxybenzoate + reduced ferredoxin + O2 | - |
Rhodopseudomonas palustris | 4-hydroxybenzoate + formaldehyde + ferredoxin + H2O | - |
? | |
1.14.99.15 | 4-methoxybenzoate + reduced putidaredoxin + O2 | very low activity with putidaredoxin | Rhodopseudomonas palustris | 4-hydroxybenzoate + formaldehyde + oxidized putidaredoxin + H2O | - |
? | |
1.14.99.15 | additional information | CYP199A2 shows a strong preference for para-substituted benzoate over identically substituted ortho- and meta- benzoates, and para-substituted benzenes, benzyl alcohols and benzaldehydes, a cytochrome P450 enzyme, the substrate binding pocket is hydrophobic, with Ser97 and Ser247 being the only polar residues, substrate binding and substrate channeling mechanism and structure, overview | Rhodopseudomonas palustris | ? | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.14.99.15 | CYP199A2 | - |
Rhodopseudomonas palustris |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.14.99.15 | 30 | - |
assay at | Rhodopseudomonas palustris |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.14.99.15 | 7.4 | - |
assay at | Rhodopseudomonas palustris |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.14.99.15 | reduced ferredoxin | - |
Rhodopseudomonas palustris |