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Literature summary for 2.3.1.233 extracted from
- Molecular analysis of the role of tyrosine 224 in the active site of Streptomyces coelicolor RppA, a bacterial type III polyketide synthase (2007), J. Biol. Chem., 282, 12765-12772.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21(DE3) cells | Streptomyces coelicolor |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C138A | inactive with malonyl-CoA | Streptomyces coelicolor |
| C138S | inactive with malonyl-CoA | Streptomyces coelicolor |
| Y224A | the mutant shows 3% activity with malonyl-CoA compared to the wild type enzyme | Streptomyces coelicolor |
| Y224C | the mutant shows 50% activity with malonyl-CoA compared to the wild type enzyme | Streptomyces coelicolor |
| Y224F | the mutant shows 70% activity with malonyl-CoA compared to the wild type enzyme | Streptomyces coelicolor |
| Y224G | inactive with malonyl-CoA but capable of accepting acetoacetyl-CoA and acetyl-CoA | Streptomyces coelicolor |
| Y224H | inactive with malonyl-CoA but capable of accepting acetoacetyl-CoA and acetyl-CoA | Streptomyces coelicolor |
| Y224L | the mutant shows 65% activity with malonyl-CoA compared to the wild type enzyme | Streptomyces coelicolor |
| Y224L | the mutant shows strongly reduced catalytic efficiency with malonyl-CoA compared to the wild type enzyme | Streptomyces griseus |
| Y224M | the mutant shows 15% activity with malonyl-CoA compared to the wild type enzyme | Streptomyces coelicolor |
| Y224S | inactive with malonyl-CoA but capable of accepting acetoacetyl-CoA and acetyl-CoA | Streptomyces coelicolor |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0011 | - |
malonyl-CoA | wild type enzyme, at pH 7.4 and 30°C | Streptomyces griseus |  |
| 0.0039 | - |
malonyl-CoA | wild type enzyme, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 0.0095 | - |
malonyl-CoA | mutant enzyme Y224F, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 0.0111 | - |
malonyl-CoA | mutant enzyme Y224L, at pH 7.4 and 30°C | Streptomyces griseus | |
| 0.012 | - |
malonyl-CoA | mutant enzyme Y224C, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 0.0171 | - |
malonyl-CoA | mutant enzyme Y224M, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 0.0234 | - |
malonyl-CoA | mutant enzyme Y224L, at pH 7.4 and 30°C | Streptomyces coelicolor | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 43000 | - |
x * 43000, SDS-PAGE | Streptomyces coelicolor |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 5 malonyl-CoA | Streptomyces coelicolor | - |
1,3,6,8-tetrahydroxynaphthalene + 5 CoA + 5 CO2 + H2O | - |
? | |
| 5 malonyl-CoA | Streptomyces griseus | - |
1,3,6,8-tetrahydroxynaphthalene + 5 CoA + 5 CO2 + H2O | - |
? | |
| 5 malonyl-CoA | Streptomyces coelicolor A3 (2) | - |
1,3,6,8-tetrahydroxynaphthalene + 5 CoA + 5 CO2 + H2O | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Streptomyces coelicolor | - |
- |
- |
| Streptomyces coelicolor A3 (2) | - |
- |
- |
| Streptomyces griseus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| nickel-nitrilotriacetic acid-agarose column chromatography | Streptomyces coelicolor |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 5 malonyl-CoA | - |
Streptomyces coelicolor | 1,3,6,8-tetrahydroxynaphthalene + 5 CoA + 5 CO2 + H2O | - |
? | |
| 5 malonyl-CoA | - |
Streptomyces griseus | 1,3,6,8-tetrahydroxynaphthalene + 5 CoA + 5 CO2 + H2O | - |
? | |
| 5 malonyl-CoA | - |
Streptomyces coelicolor A3 (2) | 1,3,6,8-tetrahydroxynaphthalene + 5 CoA + 5 CO2 + H2O | - |
? | |
| acetoacetyl-CoA | - |
Streptomyces griseus | 4-hydroxy-6-methyl-2-pyrone + CoA + CO2 + H2O | - |
? | |
| acetoacetyl-CoA + 2 malonyl-CoA | - |
Streptomyces coelicolor | 4-hydroxy-6-methyl-2-pyrone + CoA + CO2 + H2O | - |
? | |
| acetoacetyl-CoA + 2 malonyl-CoA | - |
Streptomyces coelicolor A3 (2) | 4-hydroxy-6-methyl-2-pyrone + CoA + CO2 + H2O | - |
? | |
| acetyl-CoA | - |
Streptomyces griseus | ? + CoA + CO2 + H2O | - |
? | |
| acetyl-CoA + 2 malonyl-CoA | - |
Streptomyces coelicolor | ? + CoA + CO2 + H2O | - |
? | |
| acetyl-CoA + 2 malonyl-CoA | - |
Streptomyces coelicolor A3 (2) | ? + CoA + CO2 + H2O | - |
? | |
| benzoyl-CoA | - |
Streptomyces griseus | ? + CoA + CO2 + H2O | - |
? | |
| benzoyl-CoA + 2 malonyl-CoA | - |
Streptomyces coelicolor | 4-hydroxy-6-phenyl-2H-pyran-2-one + CoA + CO2 + H2O | - |
? | |
| benzoyl-CoA + 2 malonyl-CoA | - |
Streptomyces coelicolor A3 (2) | 4-hydroxy-6-phenyl-2H-pyran-2-one + CoA + CO2 + H2O | - |
? | |
| hexanoyl-CoA | - |
Streptomyces griseus | ? + CoA + CO2 + H2O | - |
? | |
| hexanoyl-CoA + 2 malonyl-CoA | - |
Streptomyces coelicolor | 4-hydroxy-6-pentyl-2H-pyran-2-one + CoA + CO2 + H2O | - |
? | |
| hexanoyl-CoA + 2 malonyl-CoA | - |
Streptomyces coelicolor A3 (2) | 4-hydroxy-6-pentyl-2H-pyran-2-one + CoA + CO2 + H2O | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 43000, SDS-PAGE | Streptomyces coelicolor |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| RppA | - |
Streptomyces coelicolor |
| RppA | - |
Streptomyces griseus |
| THN synthase | - |
Streptomyces coelicolor |
| THN synthase | - |
Streptomyces griseus |
| THNS | - |
Streptomyces coelicolor |
| THNS | - |
Streptomyces griseus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0003 | - |
malonyl-CoA | mutant enzyme Y224L, at pH 7.4 and 30°C | Streptomyces griseus | |
| 0.002 | - |
malonyl-CoA | mutant enzyme Y224M, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 0.004 | - |
malonyl-CoA | mutant enzyme Y224L, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 0.005 | - |
malonyl-CoA | mutant enzyme Y224C, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 0.005 | - |
malonyl-CoA | wild type enzyme, at pH 7.4 and 30°C | Streptomyces griseus | |
| 0.018 | - |
malonyl-CoA | mutant enzyme Y224F, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 0.022 | - |
malonyl-CoA | wild type enzyme, at pH 7.4 and 30°C | Streptomyces coelicolor | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.133 | - |
malonyl-CoA | mutant enzyme Y224M, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 0.175 | - |
malonyl-CoA | mutant enzyme Y224L, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 0.298 | - |
malonyl-CoA | mutant enzyme Y224L, at pH 7.4 and 30°C | Streptomyces griseus | |
| 0.396 | - |
malonyl-CoA | mutant enzyme Y224C, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 1.93 | - |
malonyl-CoA | mutant enzyme Y224F, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 5.74 | - |
malonyl-CoA | wild type enzyme, at pH 7.4 and 30°C | Streptomyces coelicolor | |
| 44.58 | - |
malonyl-CoA | wild type enzyme, at pH 7.4 and 30°C | Streptomyces griseus | |
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