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Literature summary extracted from
- The role of arginine residues in substrate binding and catalysis by deacetoxycephalosporin C synthase (2002), Eur. J. Biochem., 269, 2735-2739.
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.14.20.1 | R160Q | more than 95% loss of activity | Streptomyces clavuligerus |
| 1.14.20.1 | R266L | 2-oxoglutarate conversion is very low and the same whether penicillin N, penicillin G or no penicillin substrate is present | Streptomyces clavuligerus |
| 1.14.20.1 | R266Q | 2-oxoglutarate conversion is very low and the same whether penicillin N, penicillin G or no penicillin substrate is present | Streptomyces clavuligerus |
| 1.14.20.1 | R306L | mutation enhances penicillin N conversion compared to the level of wild-type enzyme, turnover number for penicillin N is increased, no enhancement in activity with penicillin G as substrate, little effect on kinetic values using penicillin G as substrate | Streptomyces clavuligerus |
| 1.14.20.1 | R307Q | mutation enhances penicillin N conversion compared to the level of wild-type enzyme, turnover number for penicillin N is increased, no enhancement in activity with penicillin G as substrate. Mutation increase the Km-value by 10fold, but has little effect on the turnover number for penicillin G | Streptomyces clavuligerus |
| 1.14.20.1 | R74I | turnover number and Km-values are similar to that for wild-type enzyme. 2-Oxoglutarate conversion is significantly stimulated in presence of penicillin N and penicillin G compared to wild type enzyme. Penicillin oxidation is reduced relative to the wild type enzyme | Streptomyces clavuligerus |
| 1.14.20.1 | R74Q | turnover number and Km-values are similar to that for wild-type enzyme. 2-Oxoglutarate conversion is significantly stimulated in presence of penicillin N and penicillin G compared to wild type enzyme. Penicillin oxidation is reduced relative to the wild type enzyme | Streptomyces clavuligerus |
| 1.14.20.1 | R74Q/R266I | 2-oxoglutarate conversion is not stimulated by penicillin substrates | Streptomyces clavuligerus |
| 1.14.20.1 | R74Q/R266Q | 2-oxoglutarate conversion is not stimulated by penicillin substrates | Streptomyces clavuligerus |
| 1.14.20.1 | R75I/D270G | 2-oxoglutarate conversion is not stimulated by penicillin substrates | Streptomyces clavuligerus |
| 1.14.20.1 | R75Q | turnover number and Km-values are similar to that for wild-type enzyme. 2-Oxoglutarate conversion is significantly stimulated in presence of penicillin N compared to wild type enzyme | Streptomyces clavuligerus |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.20.1 | additional information | - |
additional information | Km-values for mutant enzymes with penicillin N and penicillin G as substrate | Streptomyces clavuligerus | |
| 1.14.20.1 | 0.033 | - |
Penicillin N | wild-type enzyme | Streptomyces clavuligerus |  |
| 1.14.20.1 | 0.7 | - |
penicillin G | wild-type enzyme | Streptomyces clavuligerus | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.20.1 | penicillin N + 2-oxoglutarate + O2 | Streptomyces clavuligerus | committed step in biosynthesis of cephamycin C | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.20.1 | Streptomyces clavuligerus | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.20.1 | wild-type and mutant enzymes R74I, R74Q, R75I/D270G, R75Q, R306L and R307Q | Streptomyces clavuligerus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.20.1 | penicillin G + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | phenylacetyl-7-aminodeacetoxy cephalosporanic acid + succinate + CO2 + H2O | - |
? | |
| 1.14.20.1 | penicillin N + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
| 1.14.20.1 | penicillin N + 2-oxoglutarate + O2 | committed step in biosynthesis of cephamycin C | Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.20.1 | 0.02 | - |
Penicillin N | wild-type enzyme | Streptomyces clavuligerus | |
| 1.14.20.1 | 0.05 | - |
penicillin G | wild-type enzyme | Streptomyces clavuligerus | |
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