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Literature summary for 1.14.11.21 extracted from
- Site-directed mutagenesis and biochemical analysis of the endogenous ligands in the ferrous active site of clavaminate synthase. The His-3 variant of the 2-His-1-carboxylate model (2000), Biochemistry, 39, 8666-8673.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene cs2, isozyme CS2, wild-type and mutants, overexpression in Escherichia coli JM101 | Streptomyces clavuligerus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| H109Q | isozyme CS2, site-directed mutagensis, 83% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity | Streptomyces clavuligerus |
| H122Q | isozyme CS2, site-directed mutagensis, 96% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity | Streptomyces clavuligerus |
| H131Q | isozyme CS2, site-directed mutagensis, 16% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity | Streptomyces clavuligerus |
| H145Q | isozyme CS2, site-directed mutagensis, no activity of clavaminate formation, but slight hydroxylating activity of deoxyguanidinoproclavaminate | Streptomyces clavuligerus |
| H151Q | isozyme CS2, site-directed mutagensis, 10% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity | Streptomyces clavuligerus |
| H167Q | isozyme CS2, site-directed mutagensis, 95% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity | Streptomyces clavuligerus |
| H280Q | isozyme CS2, site-directed mutagensis, no activity of clavaminate formation, but slight hydroxylating activity of deoxyguanidinoproclavaminate | Streptomyces clavuligerus |
| H297Q | isozyme CS2, site-directed mutagensis, 65% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity | Streptomyces clavuligerus |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Fe2+ | required | Streptomyces clavuligerus |  |
| Fe2+ | isozyme CS2, a single ferrous active site | Streptomyces clavuligerus | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Streptomyces clavuligerus | - |
isozyme CS2 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and mutants of isozyme CS2 from Escherichia coli JM101 | Streptomyces clavuligerus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| deoxyguanidinoproclavaminate + 2-oxoglutarate + O2 | hydroxylation | Streptomyces clavuligerus | guanidinoproclavaminate + succinate + CO2 + H2O | - |
? | |
| dihydroclavaminate + 2-oxoglutarate + O2 | cyclization | Streptomyces clavuligerus | clavaminate + succinate + CO2 + H2O | - |
? | |
| proclavaminate + 2-oxoglutarate + O2 | saturation | Streptomyces clavuligerus | dihydroclavaminate + succinate + CO2 + H2O | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CS | - |
Streptomyces clavuligerus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 23 | - |
assay at | Streptomyces clavuligerus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
assay at | Streptomyces clavuligerus |
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