Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Streptomyces clavuligerus |
Protein Variants | Comment | Organism |
---|---|---|
A106T | 80% relative activity with 1 mM substrate and 170% relative activity with 10 mM penicillin G compared to wild-type activity detected with 1 mM penicillin G, 1.5fold reduction in Km, penicillin G as substrate | Streptomyces clavuligerus |
A106T/C155Y | epPCR random mutagenesis | Streptomyces clavuligerus |
A106T/M188V | epPCR random mutagenesis | Streptomyces clavuligerus |
A11V/T91A/C281Y/I305L | 220% relative activity with 1 mM and 200% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
C155Y | 90% relative activity with 1 mM and 180% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, 1.5fold reduction in Km, penicillin G as substrate | Streptomyces clavuligerus |
C155Y/Y184H/V275I/C281Y | 580% relative activity with 1 mM and 670% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
C281R | activity similar to wild-type enzyme | Streptomyces clavuligerus |
C281Y | epPCR random mutagenesis | Streptomyces clavuligerus |
D107G | activity similar to wild-type enzyme | Streptomyces clavuligerus |
D107G/L277Q | epPCR random mutagenesis | Streptomyces clavuligerus |
D53H/C281Y | epPCR random mutagenesis | Streptomyces clavuligerus |
E144K/M188I/A198T/V275I/C281Y | 160% relative activity with 1 mM and 200% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
G300V | 410% relative activity with 1 mM and 370% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
G300V | substrate inhibition, epPCR random mutagenesis | Streptomyces clavuligerus |
H244Q | 140% relative activity with 1 mM and 270% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, 1.9fold reduction in Km, penicillin G as substrate, epPCR random mutagenesis | Streptomyces clavuligerus |
I305L | epPCR random mutagenesis | Streptomyces clavuligerus |
L277Q | 270% relative activity with 1 mM and 410% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, 2.5fold reduction in Km, penicillin G as substrate | Streptomyces clavuligerus |
L277Q/I305M | 610% relative activity with 1 mM and 520% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
M188I | 90% relative activity with 1 mM and 190% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, epPCR random mutagenesis | Streptomyces clavuligerus |
M188V | 150% relative activity with 1 mM and 190% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, epPCR random mutagenesis | Streptomyces clavuligerus |
M188V/V275I/G300V | 440% relative activity with 1 mM and 310% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
M73T | 180% relative activity with 1 mM and 250% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, 3.5fold reduction in Km for penicillin N and 2.3fold reduction in Km for penicillin G, epPCR random mutagenesis | Streptomyces clavuligerus |
M73T/C281Y | 610% relative activity with 1 mM and 530% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
M73T/G300V/I305L | 640% relative activity with 1 mM and 380% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
M73T/I118V/A140V/H244Q/C281Y | 450% relative activity with 1 mM and 350% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
M73T/I277Q | 350% relative activity with 1 mM and 350% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
M73T/P145L | epPCR random mutagenesis | Streptomyces clavuligerus |
R135Q/C155Y/R179Q/M188V/I305M | 360% relative activity with 1 mM and 290% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
T91A | 110% relative activity with 1 mM and 200% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, 1.9fold reduction in Km, penicillin G as substrate, epPCR random mutagenesis | Streptomyces clavuligerus |
V275I/C281Y | 260% relative activity with 1 mM and 310% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
V275I/C281Y/G300V | 620% relative activity with 1 mM and 380% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
V275I/I305L | 310% relative activity with 1 mM and 330% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
Y184H | 200% relative activity with 1 mM and 330% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
Y184H/C281R | epPCR random mutagenesis | Streptomyces clavuligerus |
Y184H/H244Q/T259I/L277Q | 320% relative activity with 1 mM and 310% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
Y184H/M188I/C281Y/I305L | 610% relative activity with 1 mM and 460% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.006 | - |
Penicillin N | mutant M73T | Streptomyces clavuligerus | |
0.009 | - |
Penicillin N | mutant H244Q | Streptomyces clavuligerus | |
0.009 | - |
Penicillin N | mutant T91A | Streptomyces clavuligerus | |
0.011 | - |
Penicillin N | mutant L277Q | Streptomyces clavuligerus | |
0.011 | - |
Penicillin N | mutant M188I | Streptomyces clavuligerus | |
0.013 | - |
Penicillin N | mutant A106T | Streptomyces clavuligerus | |
0.014 | - |
Penicillin N | wild-type | Streptomyces clavuligerus | |
0.016 | - |
Penicillin N | mutant C155Y | Streptomyces clavuligerus | |
0.047 | - |
Penicillin N | mutant M188V | Streptomyces clavuligerus | |
0.092 | - |
Penicillin N | mutant C155Y/Y184H/V275I/C281Y | Streptomyces clavuligerus | |
0.19 | - |
penicillin G | mutant C155Y/Y184H/V275I/C281Y | Streptomyces clavuligerus | |
0.295 | - |
Penicillin N | mutant Y184H | Streptomyces clavuligerus | |
0.74 | - |
penicillin G | mutant M73T | Streptomyces clavuligerus | |
0.76 | - |
penicillin G | mutant M188V | Streptomyces clavuligerus | |
0.95 | - |
penicillin G | mutant Y184H | Streptomyces clavuligerus | |
1.02 | - |
penicillin G | mutant L277Q | Streptomyces clavuligerus | |
1.36 | - |
penicillin G | mutant T91A | Streptomyces clavuligerus | |
1.39 | - |
penicillin G | mutant H244Q | Streptomyces clavuligerus | |
1.76 | - |
penicillin G | mutant C155Y | Streptomyces clavuligerus | |
1.77 | - |
penicillin G | mutant A106T | Streptomyces clavuligerus | |
1.96 | - |
penicillin G | mutant M188I | Streptomyces clavuligerus | |
2.58 | - |
penicillin G | wild-type | Streptomyces clavuligerus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
penicillin G + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces clavuligerus | P18548 | - |
- |
Purification (Comment) | Organism |
---|---|
anion-exchange and gel filtration | Streptomyces clavuligerus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
160% relative activity with 10 mM substrate penicillin G compared to the activity detected with 1 mM penicillin G | Streptomyces clavuligerus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
penicillin G + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
penicillin N + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
DAOCS | - |
Streptomyces clavuligerus |