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Literature summary extracted from
- Purification and characterization of carbazole 1,9a-dioxygenase, a three-component dioxygenase system of Pseudomonas resinovorans strain CA10 (2002), Appl. Environ. Microbiol., 68, 5882-5890.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.14.12.22 | expression of ferredoxin subunit CarAc in Escherichia coli, His-tagged | Pseudomonas resinovorans |
| 1.14.12.22 | expression of terminal oxygenase CarAa in Escherichia coli in native form | Pseudomonas resinovorans |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.12.22 | 0.0034 | - |
NADH | CarAd activity, electron acceptor 2,6-dichlorophenolindophenol, pH 7.5, 30°C | Pseudomonas resinovorans |  |
| 1.14.12.22 | 0.182 | - |
NADPH | CarAd activity, electron acceptor 2,6-dichlorophenolindophenol, pH 7.5, 30°C | Pseudomonas resinovorans | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.12.22 | Iron | subunit CarAc, one Rieske type [2Fe-2S] cluster per monomer protein. The iron and sulfur contents of His-tagged CarAc are 1.7 to 1.9 and 1.3 to 1.6 mol/mol of protein, respectively | Pseudomonas resinovorans | |
| 1.14.12.22 | Iron | subunit CarAd, one Rieske type [2Fe-2S] cluster per monomer protein. The iron and sulfur contents of His-tagged CarAc are 1.8 to 2.0 and 1.8 to 1.9 mol/mol of protein, respectively | Pseudomonas resinovorans | |
| 1.14.12.22 | Iron | terminal oxygenase subunit contains one Rieske type [2Fe2S] cluster and one mononuclear iron center in each monomer | Pseudomonas resinovorans | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.14.12.22 | 13000 | - |
gel filtration | Pseudomonas resinovorans |
| 1.14.12.22 | 37000 | - |
gel filtration | Pseudomonas resinovorans |
| 1.14.12.22 | 43782 | - |
3 * 44000, SDS-PAGE, 3 * 43782, calculated, subunit CarAa | Pseudomonas resinovorans |
| 1.14.12.22 | 44000 | - |
3 * 44000, SDS-PAGE, 3 * 43782, calculated, subunit CarAa | Pseudomonas resinovorans |
| 1.14.12.22 | 132000 | - |
gel filtration, subunit CarAa | Pseudomonas resinovorans |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.12.22 | Pseudomonas resinovorans | - |
- |
- |
| 1.14.12.22 | Pseudomonas resinovorans | Q8GI14 | ferredoxin reductase subunit CarAd | - |
| 1.14.12.22 | Pseudomonas resinovorans | Q8GI16 | ferredoxin subunit CarAc | - |
| 1.14.12.22 | Pseudomonas resinovorans CA10 | - |
- |
- |
| 1.14.12.22 | Pseudomonas resinovorans CA10 | Q8GI14 | ferredoxin reductase subunit CarAd | - |
| 1.14.12.22 | Pseudomonas resinovorans CA10 | Q8GI16 | ferredoxin subunit CarAc | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.12.22 | recombinant protein | Pseudomonas resinovorans |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 1.14.12.22 | 0°C 10% glycerol, 8 days, more than 90% residual activity, isolated subunit CarAa | Pseudomonas resinovorans |
| 1.14.12.22 | 0°C 50 mM Tris-HCl pH 7.5, 24 h, 90% residual activity, reconstitued CARDO system | Pseudomonas resinovorans |
| 1.14.12.22 | 0°C, 10 h, full activity, and 100 h, 93% residual activiy, ferredoxin reductase subunit CarAd | Pseudomonas resinovorans |
| 1.14.12.22 | 0°C, 10% glycerol, 8 days, more than 90% residual activity for isolated subunit CarAa | Pseudomonas resinovorans |
| 1.14.12.22 | 0°C, 50 mM Tris-HCl pH 7.5, 24 h, 90% residual activity for reconstituted CARDO system | Pseudomonas resinovorans |
| 1.14.12.22 | 4°C, 10% glycerol, 24 h, full activity for isolated subunit CarAa | Pseudomonas resinovorans |
| 1.14.12.22 | 4°C, 10% glycerol, 24 h, full activity, isolated subunit CarAa | Pseudomonas resinovorans |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.12.22 | carbazole + NADH + H+ + O2 | - |
Pseudomonas resinovorans | 2'-aminobiphenyl-2,3-diol + NAD+ | - |
? | |
| 1.14.12.22 | carbazole + NADH + H+ + O2 | - |
Pseudomonas resinovorans CA10 | 2'-aminobiphenyl-2,3-diol + NAD+ | - |
? | |
| 1.14.12.22 | carbazole + NADPH + H+ + O2 | - |
Pseudomonas resinovorans | 2'-aminobiphenyl-2,3-diol + NADP+ | - |
? | |
| 1.14.12.22 | carbazole + NADPH + H+ + O2 | - |
Pseudomonas resinovorans CA10 | 2'-aminobiphenyl-2,3-diol + NADP+ | - |
? | |
| 1.14.12.22 | additional information | in the presence of NADH, His-tagged ferrdoxin subunit CarAc is reduced by His-tagged ferredoxin reductase CarAd. Terminal oxygenase subunit CarAa is reduced by His-tagged CarAc, His-tagged CarAd, and NADH. The three purified proteins CarAa, CarAc and CarAd can reconstitute the CARDO activity in vitro. In the reconstituted CARDO system, His-tagged CarAc is indispensable for electron transport, while His-tagged CarAd can be replaced by some unrelated reductases | Pseudomonas resinovorans | ? | - |
? | |
| 1.14.12.22 | additional information | in the presence of NADH, His-tagged ferrdoxin subunit CarAc is reduced by His-tagged ferredoxin reductase CarAd. The three purified proteins CarAa, CarAc and CarAd can reconstitute the CARDO activity in vitro. In the reconstituted CARDO system, His-tagged CarAc is indispensable for electron transport, while His-tagged CarAd can be replaced by some unrelated reductases | Pseudomonas resinovorans | ? | - |
? | |
| 1.14.12.22 | additional information | terminal oxygenase subunit CarAa is reduced by His-tagged ferredoxin CarAc, His-tagged ferredoxin reductase CarAd, and NADH. The three purified proteins can reconstitute the CARDO activity in vitro. In the reconstituted CARDO system, His-tagged CarAc is indispensable for electron transport, while His-tagged CarAd can be replaced by some unrelated reductases | Pseudomonas resinovorans | ? | - |
? | |
| 1.14.12.22 | additional information | terminal oxygenase subunit CarAa is reduced by His-tagged ferredoxin CarAc, His-tagged ferredoxin reductase CarAd, and NADH. The three purified proteins can reconstitute the CARDO activity in vitro. In the reconstituted CARDO system, His-tagged CarAc is indispensable for electron transport, while His-tagged CarAd can be replaced by some unrelated reductases | Pseudomonas resinovorans CA10 | ? | - |
? | |
| 1.14.12.22 | additional information | in the presence of NADH, His-tagged ferrdoxin subunit CarAc is reduced by His-tagged ferredoxin reductase CarAd. Terminal oxygenase subunit CarAa is reduced by His-tagged CarAc, His-tagged CarAd, and NADH. The three purified proteins CarAa, CarAc and CarAd can reconstitute the CARDO activity in vitro. In the reconstituted CARDO system, His-tagged CarAc is indispensable for electron transport, while His-tagged CarAd can be replaced by some unrelated reductases | Pseudomonas resinovorans CA10 | ? | - |
? | |
| 1.14.12.22 | additional information | in the presence of NADH, His-tagged ferrdoxin subunit CarAc is reduced by His-tagged ferredoxin reductase CarAd. The three purified proteins CarAa, CarAc and CarAd can reconstitute the CARDO activity in vitro. In the reconstituted CARDO system, His-tagged CarAc is indispensable for electron transport, while His-tagged CarAd can be replaced by some unrelated reductases | Pseudomonas resinovorans CA10 | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.14.12.22 | monomer | 1 * 13000, SDS-PAGE, subunit CarAc | Pseudomonas resinovorans |
| 1.14.12.22 | monomer | 1 * 37000, SDS-PAGE, subunit CarAd | Pseudomonas resinovorans |
| 1.14.12.22 | trimer | 3 * 44000, SDS-PAGE, 3 * 43782, calculated, subunit CarAa | Pseudomonas resinovorans |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.12.22 | 30 | - |
- |
Pseudomonas resinovorans |
| 1.14.12.22 | 35 | 45 | maximum cytochrome c reductase activity of subunit CarAd | Pseudomonas resinovorans |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.12.22 | 7.5 | - |
- |
Pseudomonas resinovorans |
| 1.14.12.22 | 7.5 | 8.5 | maximum cytochrome c reductase activity of subunit CarAd | Pseudomonas resinovorans |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.12.22 | FAD | 1 mol of His-tagged CarAd contains 1 mol of FAD | Pseudomonas resinovorans | |
| 1.14.12.22 | NADH | both NADH and NADPH are effective as electron donors for His-tagged ferrdoxin reductase CarAd. The ratio kcat/Km for NADH is 22.3-fold higher than that for NADPH in the 2,6-dichlorophenolindophenol reductase assay | Pseudomonas resinovorans | |
| 1.14.12.22 | NADPH | both NADH and NADPH are effective as electron donors for His-tagged ferrdoxin reductase CarAd. The ratio kcat/Km for NADH is 22.3-fold higher than that for NADPH in the 2,6-dichlorophenolindophenol reductase assay | Pseudomonas resinovorans | |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.12.22 | 130 | - |
NADPH | CarAd activity, electron acceptor 2,6-dichlorophenolindophenol, pH 7.5, 30°C | Pseudomonas resinovorans | |
| 1.14.12.22 | 2900 | - |
NADH | CarAd activity, electron acceptor 2,6-dichlorophenolindophenol, pH 7.5, 30°C | Pseudomonas resinovorans | |
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