EC Number | General Stability | Organism |
---|---|---|
1.2.7.1 | following dialysis, the enzyme is very unstable in the absence of glycerol or ethylene glycol, thiamine diphosphate and MgCl2 also help to maintain activity | Methanococcus maripaludis |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
1.2.7.1 | Dithionite | with 5 mM dithionite, the half-life is 7 h at 2 °C, with about 90% of the original activity being lost after 24 h | Methanococcus maripaludis | |
1.2.7.1 | glyoxylate | 20 mM, about 40% of the original activity is lost. 2 mM glyoxylate inhibits 6% | Methanococcus maripaludis | |
1.2.7.1 | additional information | no substrate inhibition with CoA up to 0.1 mM. Slightly or not inhibited at all by glyoxylate, nitrite, CO or potential physiological effectors. Not inhibited by CO. Potential affectors such as ATP, ADP, AMP, cAMP, GTP, GDP, GMP, NAD+, NADH, and glyceraldehyde 3-phosphate, at concentrations of 2 mM, do not affect the activity | Methanococcus maripaludis | |
1.2.7.1 | oxygen | - |
Methanococcus maripaludis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.2.7.1 | 0.0058 | - |
CoA | pH 8.6, 37°C | Methanococcus maripaludis | |
1.2.7.1 | 0.115 | - |
pyruvate | pH 8.6, 37°C | Methanococcus maripaludis | |
1.2.7.1 | 0.205 | - |
2-oxobutyrate | pH 8.6, 37°C | Methanococcus maripaludis | |
1.2.7.1 | 0.264 | - |
oxaloacetate | pH 8.6, 37°C | Methanococcus maripaludis |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.2.7.1 | Iron-sulfur cluster | porE encodes the 21500 Da subunit that contains a high cysteinyl residue content and a motif indicative of a [FeS] cluster. Subunit porF also also has a high cysteinyl residue content, and two [FeS] cluster motifs. Based upon these results, it is proposed that PorE and PorF are components of a specialized system required to transfer low-potential electrons for pyruvate biosynthesis | Methanococcus maripaludis |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.2.7.1 | 190000 | - |
- |
Methanococcus maripaludis |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.2.7.1 | Methanococcus maripaludis | Q9P9E5 and Q9P9E4 and Q9P9E6 and Q9P9E7 and Q9P9E3 | Q9P9E5: subunit alpha, Q9P9E4: subunit beta, Q9P9E6: subunit gamma, Q9P9E7: subunit delta, Q9P9E3: subunit PorE | - |
1.2.7.1 | Methanococcus maripaludis DSM Z 2067 | Q9P9E5 and Q9P9E4 and Q9P9E6 and Q9P9E7 and Q9P9E3 | Q9P9E5: subunit alpha, Q9P9E4: subunit beta, Q9P9E6: subunit gamma, Q9P9E7: subunit delta, Q9P9E3: subunit PorE | - |
EC Number | Oxidation Stability | Organism |
---|---|---|
1.2.7.1 | the enzyme is very sensitive to O2. Following incubation in air at 2°C for 40 min, about 60% of enzyme activity is lost. The half-life is 5.2 min when the purified enzyme is exposed to air in an ice bath. After inactivation of the purified enzyme by oxygen, activity is not restored byreplacing the oxygen with nitrogen and adding 0.01 mM dithionite no activity is lost after dialysis of extract in a basic buffer containing 20 mM potassium Tricine, pH 8.6, 5 mM MgCl2, 0.5 mM dithiothreitol, 0.1 mM thiamine diphosphate, and 10% glycerol | Methanococcus maripaludis |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.2.7.1 | - |
Methanococcus maripaludis |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
1.2.7.1 | 0.3 | - |
pH 8.6, 37°C, substrate: indol-3-pyruvate | Methanococcus maripaludis |
1.2.7.1 | 3.6 | - |
pH 8.6, 37°C, substrate: 2-oxobutyrate | Methanococcus maripaludis |
1.2.7.1 | 6.5 | - |
pH 8.6, 37°C, substrate: oxaloacetate | Methanococcus maripaludis |
1.2.7.1 | 7.4 | - |
pH 8.6, 37°C, substrate: pyruvate | Methanococcus maripaludis |
EC Number | Storage Stability | Organism |
---|---|---|
1.2.7.1 | -20°C, 3 weeks, anaerobic conditions, no loss of activity of the partially purified enzyme | Methanococcus maripaludis |
1.2.7.1 | 2°C, 2 weeks, anaerobic conditions, no loss of activity | Methanococcus maripaludis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.2.7.1 | 2-oxobutyrate + CoA + oxidized methyl viologen | the enzyme has a broad substrate specificity. In the presence of CoA, it oxidizes pyruvate, oxaloacetate, 2-oxobutyrate, and indol-3-pyruvate with specific activities of 7.4, 6.5, 3.6, and 0.3 U/mg, respectively. The enzyme reduces clostridial rubredoxin, clostridial and spinach ferredoxin, cytochrome c, FMN, and FAD. No activity is detected with NAD+, NADP+, and vitamin K1. The catalytic efficiencies or kcat/Km values for FAD and FMN are calculated to be 2400032000/min * M, which are about two orders of magnitude lower than observed for the likely physiological electron carriers of other pyruvate oxidoreductases. Therefore, it is unlikely that flavins are the physiological electron carrier of the methanococcal pyruvate oxidoreductase | Methanococcus maripaludis | propanoyl-CoA + CO2 + reduced methyl viologen | - |
? | |
1.2.7.1 | 2-oxobutyrate + CoA + oxidized methyl viologen | the enzyme has a broad substrate specificity. In the presence of CoA, it oxidizes pyruvate, oxaloacetate, 2-oxobutyrate, and indol-3-pyruvate with specific activities of 7.4, 6.5, 3.6, and 0.3 U/mg, respectively. The enzyme reduces clostridial rubredoxin, clostridial and spinach ferredoxin, cytochrome c, FMN, and FAD. No activity is detected with NAD+, NADP+, and vitamin K1. The catalytic efficiencies or kcat/Km values for FAD and FMN are calculated to be 2400032000/min * M, which are about two orders of magnitude lower than observed for the likely physiological electron carriers of other pyruvate oxidoreductases. Therefore, it is unlikely that flavins are the physiological electron carrier of the methanococcal pyruvate oxidoreductase | Methanococcus maripaludis DSM Z 2067 | propanoyl-CoA + CO2 + reduced methyl viologen | - |
? | |
1.2.7.1 | indol-3 pyruvate + CoA + 2 oxidized methyl viologen | the enzyme has a broad substrate specificity. In the presence of CoA, it oxidizes pyruvate, oxaloacetate, 2-oxobutyrate, and indol-3-pyruvate with specific activities of 7.4, 6.5, 3.6, and 0.3 U/mg, respectively. The enzyme reduces clostridial rubredoxin, clostridial and spinach ferredoxin, cytochrome c, FMN, and FAD. No activity is detected with NAD+, NADP+, and vitamin K1. The catalytic efficiencies or kcat/Km values for FAD and FMN are calculated to be 2400032000/min * M, which are about two orders of magnitude lower than observed for the likely physiological electron carriers of other pyruvate oxidoreductases. Therefore, it is unlikely that flavins are the physiological electron carrier of the methanococcal pyruvate oxidoreductase | Methanococcus maripaludis | ? | - |
? | |
1.2.7.1 | indol-3 pyruvate + CoA + 2 oxidized methyl viologen | the enzyme has a broad substrate specificity. In the presence of CoA, it oxidizes pyruvate, oxaloacetate, 2-oxobutyrate, and indol-3-pyruvate with specific activities of 7.4, 6.5, 3.6, and 0.3 U/mg, respectively. The enzyme reduces clostridial rubredoxin, clostridial and spinach ferredoxin, cytochrome c, FMN, and FAD. No activity is detected with NAD+, NADP+, and vitamin K1. The catalytic efficiencies or kcat/Km values for FAD and FMN are calculated to be 2400032000/min * M, which are about two orders of magnitude lower than observed for the likely physiological electron carriers of other pyruvate oxidoreductases. Therefore, it is unlikely that flavins are the physiological electron carrier of the methanococcal pyruvate oxidoreductase | Methanococcus maripaludis DSM Z 2067 | ? | - |
? | |
1.2.7.1 | oxaloacetate + CoA + oxidized methyl viologen | the enzyme has a broad substrate specificity. In the presence of CoA, it oxidizes pyruvate, oxaloacetate, 2-oxobutyrate, and indol-3-pyruvate with specific activities of 7.4, 6.5, 3.6, and 0.3 U/mg, respectively. The enzyme reduces clostridial rubredoxin, clostridial and spinach ferredoxin, cytochrome c, FMN, and FAD. No activity is detected with NAD+, NADP+, and vitamin K1. The catalytic efficiencies or kcat/Km values for FAD and FMN are calculated to be 2400032000/min * M, which are about two orders of magnitude lower than observed for the likely physiological electron carriers of other pyruvate oxidoreductases. Therefore, it is unlikely that flavins are the physiological electron carrier of the methanococcal pyruvate oxidoreductase | Methanococcus maripaludis | ? | - |
? | |
1.2.7.1 | oxaloacetate + CoA + oxidized methyl viologen | the enzyme has a broad substrate specificity. In the presence of CoA, it oxidizes pyruvate, oxaloacetate, 2-oxobutyrate, and indol-3-pyruvate with specific activities of 7.4, 6.5, 3.6, and 0.3 U/mg, respectively. The enzyme reduces clostridial rubredoxin, clostridial and spinach ferredoxin, cytochrome c, FMN, and FAD. No activity is detected with NAD+, NADP+, and vitamin K1. The catalytic efficiencies or kcat/Km values for FAD and FMN are calculated to be 2400032000/min * M, which are about two orders of magnitude lower than observed for the likely physiological electron carriers of other pyruvate oxidoreductases. Therefore, it is unlikely that flavins are the physiological electron carrier of the methanococcal pyruvate oxidoreductase | Methanococcus maripaludis DSM Z 2067 | ? | - |
? | |
1.2.7.1 | pyruvate + CoA + 2 oxidized ferredoxin | the enzyme has a broad substrate specificity. In the presence of CoA, it oxidizes pyruvate, oxaloacetate, 2-oxobutyrate, and indol-3-pyruvate with specific activities of 7.4, 6.5, 3.6, and 0.3 U/mg, respectively. The enzyme reduces clostridial rubredoxin, clostridial and spinach ferredoxin, cytochrome c, FMN, and FAD. No activity is detected with NAD+, NADP+, and vitamin K1. The catalytic efficiencies or kcat/Km values for FAD and FMN are calculated to be 2400032000/min * M, which are about two orders of magnitude lower than observed for the likely physiological electron carriers of other pyruvate oxidoreductases. Therefore, it is unlikely that flavins are the physiological electron carrier of the methanococcal pyruvate oxidoreductase | Methanococcus maripaludis | acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+ | - |
? | |
1.2.7.1 | pyruvate + CoA + 2 oxidized ferredoxin | the enzyme has a broad substrate specificity. In the presence of CoA, it oxidizes pyruvate, oxaloacetate, 2-oxobutyrate, and indol-3-pyruvate with specific activities of 7.4, 6.5, 3.6, and 0.3 U/mg, respectively. The enzyme reduces clostridial rubredoxin, clostridial and spinach ferredoxin, cytochrome c, FMN, and FAD. No activity is detected with NAD+, NADP+, and vitamin K1. The catalytic efficiencies or kcat/Km values for FAD and FMN are calculated to be 2400032000/min * M, which are about two orders of magnitude lower than observed for the likely physiological electron carriers of other pyruvate oxidoreductases. Therefore, it is unlikely that flavins are the physiological electron carrier of the methanococcal pyruvate oxidoreductase | Methanococcus maripaludis DSM Z 2067 | acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+ | - |
? | |
1.2.7.1 | pyruvate + CoA + 2 oxidized methyl viologen | the enzyme has a broad substrate specificity. In the presence of CoA, it oxidizes pyruvate, oxaloacetate, 2-oxobutyrate, and indol-3-pyruvate with specific activities of 7.4, 6.5, 3.6, and 0.3 U/mg, respectively. The enzyme reduces clostridial rubredoxin, clostridial and spinach ferredoxin, cytochrome c, FMN, and FAD. No activity is detected with NAD+, NADP+, and vitamin K1. The catalytic efficiencies or kcat/Km values for FAD and FMN are calculated to be 2400032000/min * M, which are about two orders of magnitude lower than observed for the likely physiological electron carriers of other pyruvate oxidoreductases. Therefore, it is unlikely that flavins are the physiological electron carrier of the methanococcal pyruvate oxidoreductase | Methanococcus maripaludis | acetyl-CoA + CO2 + 2 reduced methyl viologen + 2 H+ | - |
? | |
1.2.7.1 | pyruvate + CoA + 2 oxidized methyl viologen | the enzyme has a broad substrate specificity. In the presence of CoA, it oxidizes pyruvate, oxaloacetate, 2-oxobutyrate, and indol-3-pyruvate with specific activities of 7.4, 6.5, 3.6, and 0.3 U/mg, respectively. The enzyme reduces clostridial rubredoxin, clostridial and spinach ferredoxin, cytochrome c, FMN, and FAD. No activity is detected with NAD+, NADP+, and vitamin K1. The catalytic efficiencies or kcat/Km values for FAD and FMN are calculated to be 2400032000/min * M, which are about two orders of magnitude lower than observed for the likely physiological electron carriers of other pyruvate oxidoreductases. Therefore, it is unlikely that flavins are the physiological electron carrier of the methanococcal pyruvate oxidoreductase | Methanococcus maripaludis DSM Z 2067 | acetyl-CoA + CO2 + 2 reduced methyl viologen + 2 H+ | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.2.7.1 | ? | the low molecular weight enzyme contains five polypeptides: 47000 Da (alpha), 33000 Da (beta), 25000 (gamma) and 13000 Da (gamma). The subunit stoichiometry for the alpha:beta:gamma:delta subunits is 1:1.08:0.90:1.32. In addition it contains a fifth polypeptide (21500 Da) | Methanococcus maripaludis |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.2.7.1 | POR | - |
Methanococcus maripaludis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.2.7.1 | 37 | - |
assay at | Methanococcus maripaludis |
1.2.7.1 | 60 | - |
at pH 7.3, the temperature optimum is 60°C | Methanococcus maripaludis |
EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.2.7.1 | 37 | 60 | the activity detected at 60°C is five times the activity detected at 37°C | Methanococcus maripaludis |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.2.7.1 | 13 | - |
2-oxobutyrate | pH 8.6, 37°C | Methanococcus maripaludis | |
1.2.7.1 | 18 | - |
oxaloacetate | pH 8.6, 37°C | Methanococcus maripaludis | |
1.2.7.1 | 27 | - |
pyruvate | pH 8.6, 37°C | Methanococcus maripaludis |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.2.7.1 | 7.3 | - |
at 37 °C, maximal activity is obtained at pH 7.3 | Methanococcus maripaludis |
1.2.7.1 | 8.6 | - |
assay at | Methanococcus maripaludis |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.2.7.1 | FAD | it is possible that flavins play an important regulatory or structural role in the enzyme | Methanococcus maripaludis | |
1.2.7.1 | FMN | it is possible that flavins play an important regulatory or structural role in the enzyme | Methanococcus maripaludis | |
1.2.7.1 | additional information | the enzyme is not coenzyme F420-dependent | Methanococcus maripaludis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.2.7.1 | physiological function | in autotrophic methanogens, pyruvate oxidoreductase plays a key role in the assimilation of CO2 and the biosynthesis of organic carbon | Methanococcus maripaludis |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.2.7.1 | 3.6 | - |
2-oxobutyrate | pH 8.6, 37°C | Methanococcus maripaludis | |
1.2.7.1 | 68.2 | - |
oxaloacetate | pH 8.6, 37°C | Methanococcus maripaludis | |
1.2.7.1 | 234.8 | - |
pyruvate | pH 8.6, 37°C | Methanococcus maripaludis |