Crystallization (Comment) | Organism |
---|---|
crystallization of wild-type enzyme and of the DELTAR306 mutant complexed with iron(II) and 2-oxoglutarate to 2.1 A and the DELTAR306A mutant complexed with iron(II), succinate and unhydrated carbon dioxide to 1.96 A | Streptomyces clavuligerus |
Protein Variants | Comment | Organism |
---|---|---|
DELTAI305-310 | mutant truncated by six residues, reduced turnover number for penicillin N compared to wild-type enzyme, lower KM-value for penicillin N compared to wild-type enzyme | Streptomyces clavuligerus |
DELTAK310-314 | mutant truncated by five residues, increased turnover number for penicillin N compared to wild-type enzyme, lower KM-value for penicillin N compared to wild-type enzyme, lower Km-value for penicillin G compared to wild-type enzyme, slightly higher turnover number for penicillin G compared to wild-type enzyme | Streptomyces clavuligerus |
DELTAR306-310 | mutant truncated by five residues, reduced turnover number for penicillin N compared to wild-type enzyme, lower KM-value for penicillin N compared to wild-type enzyme | Streptomyces clavuligerus |
DELTAR307-310 | mutant in which residues 307-310 are excised, reduced turnover number for penicillin N compared to wild-type enzyme, higher KM-value for penicillin N compared to wild-type enzyme, increased KM-value for penicillin G compared to wild-type enzyme, increased turnover-number for penicillin G compared to wild-type enzyme | Streptomyces clavuligerus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.025 | - |
Penicillin N | mutant enzyme DELTAK310-314 | Streptomyces clavuligerus | |
0.026 | - |
Penicillin N | mutant enzyme DELTAR306-310 | Streptomyces clavuligerus | |
0.034 | - |
Penicillin N | mutant enzyme DELTAI305-310 | Streptomyces clavuligerus | |
0.036 | - |
Penicillin N | wild-type enzyme | Streptomyces clavuligerus | |
0.044 | - |
Penicillin N | mutant enzyme DELTAR307-310 | Streptomyces clavuligerus | |
0.5 | - |
penicillin G | mutant enzyme DELTAK310-314 | Streptomyces clavuligerus | |
0.7 | - |
penicillin G | wild-type enzyme | Streptomyces clavuligerus | |
1.6 | - |
penicillin G | mutant enzyme DELTAR307-310 | Streptomyces clavuligerus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
penicillin N + 2-oxoglutarate + O2 | Streptomyces clavuligerus | the enzyme catalyzes the committed step in the biosynthesis of cephalosporin antibiotics | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces clavuligerus | P18548 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme expressed in Escherichia coli | Streptomyces clavuligerus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
penicillin G + 2-oxo-4-methylpentanoate + O2 | - |
Streptomyces clavuligerus | phenylacetyl-7-aminodeacetoxy cephalosporanic acid + ? | - |
? | |
penicillin G + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | phenylacetyl-7-aminodeacetoxy cephalosporanic acid + succinate + CO2 + H2O | - |
? | |
penicillin G + 2-oxohexanoate + O2 | - |
Streptomyces clavuligerus | phenylacetyl-7-aminodeacetoxy cephalosporanic acid + ? | - |
? | |
penicillin N + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
penicillin N + 2-oxoglutarate + O2 | the enzyme catalyzes the committed step in the biosynthesis of cephalosporin antibiotics | Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
Penicillin N | mutant enzyme DELTAI305-310, DELTAR306-310 and mutant DELTAR307-310 | Streptomyces clavuligerus | |
0.05 | - |
penicillin G | wild-type enzyme | Streptomyces clavuligerus | |
0.05 | - |
Penicillin N | wild-type enzyme | Streptomyces clavuligerus | |
0.06 | - |
penicillin G | mutant enzyme DELTAK310-314 | Streptomyces clavuligerus | |
0.08 | - |
penicillin G | mutant enzyme DELTAR307-310 | Streptomyces clavuligerus | |
0.08 | - |
Penicillin N | mutant enzyme DELTAI310-314 | Streptomyces clavuligerus |