Cloned (Comment) | Organism |
---|---|
expression of ferredoxin subunit CarAc in Escherichia coli, His-tagged | Pseudomonas resinovorans |
expression of terminal oxygenase CarAa in Escherichia coli in native form | Pseudomonas resinovorans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0034 | - |
NADH | CarAd activity, electron acceptor 2,6-dichlorophenolindophenol, pH 7.5, 30°C | Pseudomonas resinovorans | |
0.182 | - |
NADPH | CarAd activity, electron acceptor 2,6-dichlorophenolindophenol, pH 7.5, 30°C | Pseudomonas resinovorans |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
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 | |
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 | |
Iron | terminal oxygenase subunit contains one Rieske type [2Fe2S] cluster and one mononuclear iron center in each monomer | Pseudomonas resinovorans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
13000 | - |
gel filtration | Pseudomonas resinovorans |
37000 | - |
gel filtration | Pseudomonas resinovorans |
43782 | - |
3 * 44000, SDS-PAGE, 3 * 43782, calculated, subunit CarAa | Pseudomonas resinovorans |
44000 | - |
3 * 44000, SDS-PAGE, 3 * 43782, calculated, subunit CarAa | Pseudomonas resinovorans |
132000 | - |
gel filtration, subunit CarAa | Pseudomonas resinovorans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas resinovorans | - |
- |
- |
Pseudomonas resinovorans | Q8GI14 | ferredoxin reductase subunit CarAd | - |
Pseudomonas resinovorans | Q8GI16 | ferredoxin subunit CarAc | - |
Pseudomonas resinovorans CA10 | - |
- |
- |
Pseudomonas resinovorans CA10 | Q8GI14 | ferredoxin reductase subunit CarAd | - |
Pseudomonas resinovorans CA10 | Q8GI16 | ferredoxin subunit CarAc | - |
Purification (Comment) | Organism |
---|---|
recombinant protein | Pseudomonas resinovorans |
Storage Stability | Organism |
---|---|
0°C 10% glycerol, 8 days, more than 90% residual activity, isolated subunit CarAa | Pseudomonas resinovorans |
0°C 50 mM Tris-HCl pH 7.5, 24 h, 90% residual activity, reconstitued CARDO system | Pseudomonas resinovorans |
0°C, 10 h, full activity, and 100 h, 93% residual activiy, ferredoxin reductase subunit CarAd | Pseudomonas resinovorans |
0°C, 10% glycerol, 8 days, more than 90% residual activity for isolated subunit CarAa | Pseudomonas resinovorans |
0°C, 50 mM Tris-HCl pH 7.5, 24 h, 90% residual activity for reconstituted CARDO system | Pseudomonas resinovorans |
4°C, 10% glycerol, 24 h, full activity for isolated subunit CarAa | Pseudomonas resinovorans |
4°C, 10% glycerol, 24 h, full activity, isolated subunit CarAa | Pseudomonas resinovorans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
carbazole + NADH + H+ + O2 | - |
Pseudomonas resinovorans | 2'-aminobiphenyl-2,3-diol + NAD+ | - |
? | |
carbazole + NADH + H+ + O2 | - |
Pseudomonas resinovorans CA10 | 2'-aminobiphenyl-2,3-diol + NAD+ | - |
? | |
carbazole + NADPH + H+ + O2 | - |
Pseudomonas resinovorans | 2'-aminobiphenyl-2,3-diol + NADP+ | - |
? | |
carbazole + NADPH + H+ + O2 | - |
Pseudomonas resinovorans CA10 | 2'-aminobiphenyl-2,3-diol + NADP+ | - |
? | |
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 | ? | - |
? | |
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 | ? | - |
? | |
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 | ? | - |
? | |
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 | ? | - |
? | |
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 | ? | - |
? | |
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 | Comment | Organism |
---|---|---|
monomer | 1 * 13000, SDS-PAGE, subunit CarAc | Pseudomonas resinovorans |
monomer | 1 * 37000, SDS-PAGE, subunit CarAd | Pseudomonas resinovorans |
trimer | 3 * 44000, SDS-PAGE, 3 * 43782, calculated, subunit CarAa | Pseudomonas resinovorans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
- |
Pseudomonas resinovorans |
35 | 45 | maximum cytochrome c reductase activity of subunit CarAd | Pseudomonas resinovorans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Pseudomonas resinovorans |
7.5 | 8.5 | maximum cytochrome c reductase activity of subunit CarAd | Pseudomonas resinovorans |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | 1 mol of His-tagged CarAd contains 1 mol of FAD | Pseudomonas resinovorans | |
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 | |
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 Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
130 | - |
NADPH | CarAd activity, electron acceptor 2,6-dichlorophenolindophenol, pH 7.5, 30°C | Pseudomonas resinovorans | |
2900 | - |
NADH | CarAd activity, electron acceptor 2,6-dichlorophenolindophenol, pH 7.5, 30°C | Pseudomonas resinovorans |