Cloned (Comment) | Organism |
---|---|
expressed as a recombinant fusion protein with an N-terminal 10x histidine-tag in Escherichia coli BL21(DE3) cells | Streptomyces viridochromogenes |
Protein Variants | Comment | Organism |
---|---|---|
D297E | catalytic turnover rate is 1.5fold less and Km is increased about 13fold compared to the wild type enzyme | Streptomyces viridochromogenes |
D297N | catalytic turnover rate of D297N is 2fold lower and Km is increased 940fold compared to the wild type enzyme | Streptomyces viridochromogenes |
G94A | compared to wild type PPDC, Km is increased only 2fold, while kcat is 4fold lower | Streptomyces viridochromogenes |
H110A | no detectable activity | Streptomyces viridochromogenes |
H111A | the mutation has no significant influence on the steady state kinetic constants compared to the wild type enzyme | Streptomyces viridochromogenes |
S25D | catalytic turnover rate is 4fold less and Km is increased 460fold compared to the wild type enzyme | Streptomyces viridochromogenes |
S25N | catalytic turnover rate is 1.5fold less and Km is increased 80fold compared to the wild type enzyme | Streptomyces viridochromogenes |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0032 | - |
3-phosphonopyruvate | wild type enzyme, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes | |
0.013 | - |
3-phosphonopyruvate | mutant enzyme H111A, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes | |
0.04 | - |
3-phosphonopyruvate | mutant enzyme D297E, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes | |
0.25 | - |
3-phosphonopyruvate | mutant enzyme S25N, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes | |
1.47 | - |
3-phosphonopyruvate | mutant enzyme S25D, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes | |
3.02 | - |
3-phosphonopyruvate | mutant enzyme D297N, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | Ca2+ can substitute for Mg2+ | Streptomyces viridochromogenes | |
Mg2+ | Mg2+ is required for activity, Ca2+ and Mn2+ can substitute for Mg2+ | Streptomyces viridochromogenes | |
Mn2+ | Mn2+ can substitute for Mg2+ | Streptomyces viridochromogenes |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
43600 | - |
2 * 43600, gel filtration | Streptomyces viridochromogenes |
46000 | - |
2 * 46000, gel filtration | Streptomyces viridochromogenes |
72100 | - |
gel filtration | Streptomyces viridochromogenes |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces viridochromogenes | O86938 | strain Tü494 | - |
Streptomyces viridochromogenes DSM 40736 / JCM 4977 / BCRC 1201 / Tue 494 / Tu 494 | O86938 | strain Tü494 | - |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography | Streptomyces viridochromogenes |
Storage Stability | Organism |
---|---|
-20°C and 4°C, half-life of 50 days | Streptomyces viridochromogenes |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-phosphonopyruvate | - |
Streptomyces viridochromogenes | 2-phosphonoacetaldehyde + CO2 | - |
? | |
3-phosphonopyruvate | - |
Streptomyces viridochromogenes DSM 40736 / JCM 4977 / BCRC 1201 / Tue 494 / Tu 494 | 2-phosphonoacetaldehyde + CO2 | - |
? | |
additional information | neither pyruvate, beta-hydroxypyruvate, nor fluoropyruvate serve as alternative substrates | Streptomyces viridochromogenes | ? | - |
? | |
additional information | neither pyruvate, beta-hydroxypyruvate, nor fluoropyruvate serve as alternative substrates | Streptomyces viridochromogenes DSM 40736 / JCM 4977 / BCRC 1201 / Tue 494 / Tu 494 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 43600, gel filtration | Streptomyces viridochromogenes |
homodimer | 2 * 46000, gel filtration | Streptomyces viridochromogenes |
Synonyms | Comment | Organism |
---|---|---|
PPDC | - |
Streptomyces viridochromogenes |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.31 | - |
3-phosphonopyruvate | mutant enzyme S25D, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes | |
0.55 | - |
3-phosphonopyruvate | mutant enzyme D297N, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes | |
0.76 | - |
3-phosphonopyruvate | mutant enzyme D297E, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes | |
0.78 | - |
3-phosphonopyruvate | mutant enzyme S25N, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes | |
1.18 | - |
3-phosphonopyruvate | wild type enzyme, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes | |
1.24 | - |
3-phosphonopyruvate | mutant enzyme H111A, at pH 8.0 (HEPES buffer) and at 30°C | Streptomyces viridochromogenes |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7.5 | 8.5 | - |
Streptomyces viridochromogenes |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
thiamine diphosphate | dependent on | Streptomyces viridochromogenes |