Cloned (Comment) | Organism |
---|---|
His-tag fused tphA1II and His-tag fused tphA2IIA3II genes expressed in Escherichia coli BL21(DE3) harboring pET-16tpa1 and pET-16tpa23 respectively, under the control of the T7 promoter | Comamonas sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | 100% inhibition of TphA2IIA3II, Fe2+ restores activity, whereas Co2+, Cu2+, Mg2+, Mn2+, or Zn2+ can not restore activtiy | Comamonas sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
NADPH | - |
Comamonas sp. | |
0.051 | - |
NADH | - |
Comamonas sp. | |
0.072 | - |
terephthalate | at 30°C | Comamonas sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | is required for activity | Comamonas sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
17000 | - |
2 * 48000, SDS-PAGE, TphA2II + 2 * 17000, SDS-PAGE, TphA3II. 2 * 48763, sequence analysis, TphA2II + 2 * 17236, sequence analysis, TphA3II | Comamonas sp. |
17236 | - |
2 * 48000, SDS-PAGE, TphA2II + 2 * 17000, SDS-PAGE, TphA3II. 2 * 48763, sequence analysis, TphA2II + 2 * 17236, sequence analysis, TphA3II | Comamonas sp. |
37000 | - |
1 * 37000, SDS-PAGE, TphA1II. 1 x 38861, sequence analysis, TphA1II | Comamonas sp. |
40000 | - |
gel filtration, TphA1II | Comamonas sp. |
48000 | - |
2 * 48000, SDS-PAGE, TphA2II + 2 * 17000, SDS-PAGE, TphA3II. 2 * 48763, sequence analysis, TphA2II + 2 * 17236, sequence analysis, TphA3II | Comamonas sp. |
48763 | - |
2 * 48000, SDS-PAGE, TphA2II + 2 * 17000, SDS-PAGE, TphA3II. 2 * 48763, sequence analysis, TphA2II + 2 * 17236, sequence analysis, TphA3II | Comamonas sp. |
130000 | - |
gel filtration and native-PAGE, TphA3II | Comamonas sp. |
140000 | - |
gel filtration and native-PAGE, TphA2II | Comamonas sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Comamonas sp. | - |
- |
- |
Comamonas sp. E6 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
to near homogeneity by Ni affinity chromatography | Comamonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | does not catalyze phthalate, isophthalate, hydrobenzoate, protocatechuate, gallate, syringate, vanillate, benzoate, and 4-chlorobenzoate | Comamonas sp. | ? | - |
? | |
additional information | does not catalyze phthalate, isophthalate, hydrobenzoate, protocatechuate, gallate, syringate, vanillate, benzoate, and 4-chlorobenzoate | Comamonas sp. E6 | ? | - |
? | |
terephthalate + NAD(P)H + O2 + H+ | TPADO is specific for terephthalate | Comamonas sp. | (1R,2S)-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylate + NAD(P)+ | - |
? | |
terephthalate + NAD(P)H + O2 + H+ | TPADO is specific for terephthalate | Comamonas sp. E6 | (1R,2S)-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylate + NAD(P)+ | - |
? | |
terephthalate + NADH + O2 + H+ | - |
Comamonas sp. | (1R,2S)-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylate + NAD+ | - |
? | |
terephthalate + NADH + O2 + H+ | - |
Comamonas sp. E6 | (1R,2S)-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylate + NAD+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterotetramer | 2 * 48000, SDS-PAGE, TphA2II + 2 * 17000, SDS-PAGE, TphA3II. 2 * 48763, sequence analysis, TphA2II + 2 * 17236, sequence analysis, TphA3II | Comamonas sp. |
monomer | 1 * 37000, SDS-PAGE, TphA1II. 1 x 38861, sequence analysis, TphA1II | Comamonas sp. |
Synonyms | Comment | Organism |
---|---|---|
TPA 1,2-dioxygenase | - |
Comamonas sp. |
TPADO | - |
Comamonas sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
- |
Comamonas sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
15 | - |
about 60% of maximum TPADO activity, whereas activity is completely lost at 50°C | Comamonas sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Comamonas sp. |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
9 | - |
about 20% of maximum TPADO activity, whereas activity is completely lost at pH 5 | Comamonas sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | TphA1II contains 0.7 mol of FAD/mol of protein, it binds one FAD molecule per monomer | Comamonas sp. | |
NADH | - |
Comamonas sp. | |
NADPH | preference for NADPH over NADH | Comamonas sp. |
General Information | Comment | Organism |
---|---|---|
physiological function | TPADO is a two-component enzyme that consists of the termial oxygenase component TphA2IIA3II and the reductase component TphA1II | Comamonas sp. |