EC Number | Cloned (Comment) | Organism |
---|---|---|
1.14.15.1 | exxpression of N-terminally His-tagged CYP101D2 in Escherichia coli strain BL21(DE3) | Novosphingobium aromaticivorans |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.14.15.1 | purified recombinant apoenzyme and camphor-bound enzyme of CYP101D2, hanging drop vapour diffusion method, mixing of 0.001 ml of 50 mg/ml protein in 20 mM Tris-HCl, pH 8.0, and 150 mM KCl, with 0.001 ml of reservoir solution containing 0.1 M Tris/HCl, pH 8.3, 2.1 M ammonium sulfate, and 4% v/v PEG 400, equilibration against 0.2 ml reservoir solution, 18°C, 1 week, for substrate-bound form soaking of crystals in camphor-containing solution, 18°C, 1 week-1 month, X-ray diffraction structure determination and analysis at 2.4 A and 2.2. A resolution, respectively, molecular replacement and structure modeling | Novosphingobium aromaticivorans |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.15.1 | (+)-camphor + reduced ferredoxin + O2 | Novosphingobium aromaticivorans | - |
(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O | - |
? | |
1.14.15.1 | (+)-camphor + reduced ferredoxin + O2 | Novosphingobium aromaticivorans DSM 12444 | - |
(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.14.15.1 | Novosphingobium aromaticivorans | Q2G8A2 | - |
- |
1.14.15.1 | Novosphingobium aromaticivorans DSM 12444 | Q2G8A2 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.14.15.1 | recombinant His-tagged CYP101D2 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, gel filtration, and anion exchange chromatography to over 95% purity | Novosphingobium aromaticivorans |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.15.1 | (+)-camphor + reduced ferredoxin + O2 | - |
Novosphingobium aromaticivorans | (+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O | - |
? | |
1.14.15.1 | (+)-camphor + reduced ferredoxin + O2 | substrate of CYP101D1 and CYP101D2 | Novosphingobium aromaticivorans | (+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O | - |
? | |
1.14.15.1 | (+)-camphor + reduced ferredoxin + O2 | - |
Novosphingobium aromaticivorans DSM 12444 | (+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O | - |
? | |
1.14.15.1 | (+)-camphor + reduced ferredoxin + O2 | substrate of CYP101D1 and CYP101D2 | Novosphingobium aromaticivorans DSM 12444 | (+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.14.15.1 | CYP101D1 | - |
Novosphingobium aromaticivorans |
1.14.15.1 | CYP101D2 | - |
Novosphingobium aromaticivorans |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.14.15.1 | Ferredoxin | the CYP101D2 likely ferredoxin-binding site on the proximal face is largely positively charged, similar to that of CYP101D1 | Novosphingobium aromaticivorans | |
1.14.15.1 | heme | - |
Novosphingobium aromaticivorans |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.14.15.1 | additional information | both the apoenzyme and the camphor-bound enzyme of CYP101D2 have open conformations with an access channel. In the active site of the camphor-bound form, the camphor carbonyl interacts with the heme-iron-bound water. The observed open structures may be conformers of the CYP101D2 enzyme that enable the substrate to enter the buried active site via a conformational selection mechanism. Two other potential camphor-binding sites exist: one located in the access channel, flanked by the B/C and F/G loops and the I helix, and the other in a cavity on the surface of the enzyme near the F helix side of the F/G loop. The second and third binding sites may be intermediate locations of substrate entry and translocation into the active site, substrate binding structure and multi-step substrate-binding mechanism, overview | Novosphingobium aromaticivorans |