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Literature summary for 1.14.15.1 extracted from
- Selecting of a cytochrome P450cam SeSaM library with 3-chloroindole and endosulfan - Identification of mutants that dehalogenate 3-chloroindole (2018), Biochim. Biophys. Acta, 1866, 68-79 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression in Escherichia coli | Pseudomonas putida |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| docking simulations for binding of 3-chloroindole, the wild-type does not accommodate 3-chloroindole in the active site, whereas all the mutants do. The mutants did not differ significantly in the Fe-N, Fe-C-2 or Fe-C-3 distances for 3-chloroindole | Pseudomonas putida |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D97F/P122L/Q183L/L244Q | mutant isolated by Sequence Saturation Mutagenesis, converts 3-chloroindole to isatin | Pseudomonas putida |
| E156G/V247F/V253G/F256S | mutant isolated by Sequence Saturation Mutagenesis, shows the highest maximal velocity in the conversion of 3-chloroindole to isatin | Pseudomonas putida |
| G120A/Y179H/G248S/D297H | mutant isolated by Sequence Saturation Mutagenesis, converts 3-chloroindole to isatin | Pseudomonas putida |
| G60S/Y75H | mutant isolated by Sequence Saturation Mutagenesis, shows highest Km/kcat values for the conversion of 3-chloroindole to isatin | Pseudomonas putida |
| G93C/K314R/L319M | mutant isolated by Sequence Saturation Mutagenesis, converts 3-chloroindole to isatin | Pseudomonas putida |
| T56A/N116H/D297N | mutant isolated by Sequence Saturation Mutagenesis, shows highest Km/kcat values for the conversion of 3-chloroindole to isatin | Pseudomonas putida |
| Y179H | mutant isolated by Sequence Saturation Mutagenesis, converts 3-chloroindole to isatin | Pseudomonas putida |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0046 | - |
3-chloroindole | mutant G120A/Y179H/G248S/D297H, pH 7.4, temperature not specified in the publication | Pseudomonas putida |  |
| 0.016 | - |
3-chloroindole | mutant T56A/N116H/D297N, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.026 | - |
3-chloroindole | mutant Y179H, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.041 | - |
3-chloroindole | mutant G60S/Y75H, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.09 | - |
3-chloroindole | mutant D97F/P122L/Q183L/L244Q, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.15 | - |
3-chloroindole | mutant G93C/K314R/L319M, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.44 | - |
3-chloroindole | mutant E156G/V247F/V253G/F256S, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas putida | P00183 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (+)-camphor + reduced putidaredoxin + O2 | - |
Pseudomonas putida | (R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O | - |
? | |
| 3-chloroindole + O2 + reduced putidaredoxin | no substrate of wild-type, substrate of mutants E156G/V247F/V253G/F256S, T56A/N116H/D297N and G60S/Y75H | Pseudomonas putida | isatin + H2O + Cl- + oxidized putidaredoxin + ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| P450cam | - |
Pseudomonas putida |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0014 | - |
3-chloroindole | mutant Y179H, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.0053 | - |
3-chloroindole | mutant G120A/Y179H/G248S/D297H, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.011 | - |
3-chloroindole | mutant D97F/P122L/Q183L/L244Q, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.027 | - |
3-chloroindole | mutant T56A/N116H/D297N, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.06 | - |
3-chloroindole | mutant G60S/Y75H, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.062 | - |
3-chloroindole | mutant G93C/K314R/L319M, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.186 | - |
3-chloroindole | mutant E156G/V247F/V253G/F256S, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.05 | - |
3-chloroindole | mutant Y179H, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.12 | - |
3-chloroindole | mutant D97F/P122L/Q183L/L244Q, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.42 | - |
3-chloroindole | mutant G93C/K314R/L319M, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 0.43 | - |
3-chloroindole | mutant E156G/V247F/V253G/F256S, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 1.15 | - |
3-chloroindole | mutant G120A/Y179H/G248S/D297H, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 1.49 | - |
3-chloroindole | mutant G60S/Y75H, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
| 1.68 | - |
3-chloroindole | mutant T56A/N116H/D297N, pH 7.4, temperature not specified in the publication | Pseudomonas putida | |
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