Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Ralstonia sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05641 | - |
NADPH | in 50 mM MES buffer (pH 6.4), at 22°C | Ralstonia sp. | |
0.05976 | - |
NADH | in 50 mM MES buffer (pH 6.4), at 22°C | Ralstonia sp. | |
0.1071 | - |
salicylate | in 50 mM MES buffer (pH 6.4), at 22°C | Ralstonia sp. | |
0.3284 | - |
2-Nitrophenol | in 50 mM MES buffer (pH 6.4), at 22°C | Ralstonia sp. | |
0.9398 | - |
2,6-dihydroxybenzoate | in 50 mM MES buffer (pH 6.4), at 22°C | Ralstonia sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Iron | the enzyme contains one Rieske-type [2Fe-2S] cluster and 7.5 g iron per mol enzyme | Ralstonia sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
18800 | - |
3 * 51000 + 3 * 18800, His6-tagged enzyme, calculated from amino acid sequence | Ralstonia sp. |
20000 | - |
3 * 52000 + 3 * 20000, His6-tagged enzyme, SDS-PAGE | Ralstonia sp. |
51000 | - |
3 * 51000 + 3 * 18800, His6-tagged enzyme, calculated from amino acid sequence | Ralstonia sp. |
52000 | - |
3 * 52000 + 3 * 20000, His6-tagged enzyme, SDS-PAGE | Ralstonia sp. |
195200 | - |
gel filtration | Ralstonia sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
salicylate + NADH + H+ + O2 | Ralstonia sp. | - |
2,5-dihydroxybenzoate + NAD+ + H2O | - |
? | |
salicylate + NADH + H+ + O2 | Ralstonia sp. U2 | - |
2,5-dihydroxybenzoate + NAD+ + H2O | - |
? | |
salicylate + NADPH + H+ + O2 | Ralstonia sp. | - |
2,5-dihydroxybenzoate + NADP+ + H2O | - |
? | |
salicylate + NADPH + H+ + O2 | Ralstonia sp. U2 | - |
2,5-dihydroxybenzoate + NADP+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ralstonia sp. | O52379 and O52380 | large oxygenase component and small oxagenase component | - |
Ralstonia sp. U2 | O52379 and O52380 | large oxygenase component and small oxagenase component | - |
Purification (Comment) | Organism |
---|---|
the three-component Fe-S protein complex (NagAaGHAb) of the enzyme is purified by HisTrap column chromatography and Sephadex G-25 gel filtration | Ralstonia sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
107.1 | - |
after 124fold purification, in 50 mM MES buffer (pH 6.4), at 22°C | Ralstonia sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,4-dihydroxybenzoate + NADH + H+ + O2 | - |
Ralstonia sp. | ? | - |
? | |
2,4-dihydroxybenzoate + NADH + H+ + O2 | - |
Ralstonia sp. U2 | ? | - |
? | |
2,6-dihydroxybenzoate + NADH + H+ + O2 | - |
Ralstonia sp. | ? | - |
? | |
2,6-dihydroxybenzoate + NADH + H+ + O2 | - |
Ralstonia sp. U2 | ? | - |
? | |
2-nitrophenol + NADH + H+ + O2 | - |
Ralstonia sp. | ? | - |
? | |
2-nitrophenol + NADH + H+ + O2 | - |
Ralstonia sp. U2 | ? | - |
? | |
salicylate + NADH + H+ + O2 | - |
Ralstonia sp. | 2,5-dihydroxybenzoate + NAD+ + H2O | - |
? | |
salicylate + NADH + H+ + O2 | - |
Ralstonia sp. U2 | 2,5-dihydroxybenzoate + NAD+ + H2O | - |
? | |
salicylate + NADPH + H+ + O2 | - |
Ralstonia sp. | 2,5-dihydroxybenzoate + NADP+ + H2O | - |
? | |
salicylate + NADPH + H+ + O2 | - |
Ralstonia sp. U2 | 2,5-dihydroxybenzoate + NADP+ + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterohexamer | 3 * 51000 + 3 * 18800, His6-tagged enzyme, calculated from amino acid sequence | Ralstonia sp. |
heterohexamer | 3 * 52000 + 3 * 20000, His6-tagged enzyme, SDS-PAGE | Ralstonia sp. |
Synonyms | Comment | Organism |
---|---|---|
NagGH | - |
Ralstonia sp. |
S5H | - |
Ralstonia sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Ralstonia sp. | |
NADPH | - |
Ralstonia sp. | |
[2Fe-2S]-center | - |
Ralstonia sp. |