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Literature summary for 2.7.1.40 extracted from
- A novel pyruvate kinase and its application in lactic acid production under oxygen deprivation in Corynebacterium glutamicum (2016), BMC Biotechnol., 16, 79 .
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| AMP | - |
Corynebacterium glutamicum |  |
| fructose 1,6-bisphosphate | FBP, allosteric regulation, Pyk2 is dependent on FBP activation | Corynebacterium glutamicum | |
| additional information | glucose-6-phosphate (G6P) has no significant effect on the S0.5 and kcat of Pyk2 | Corynebacterium glutamicum |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene pyk2, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Corynebacterium glutamicum |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | construction of single deletion mutants DELTApyk1 and DELTApyk2, and of the double deletion mutant DELTApyk1DELTApyk2, complementation of the DELTApyk1DELTApyk2 strain with the pyk2 gene | Corynebacterium glutamicum |
| additional information | construction of single deletion mutants DELTApyk1 and DELTApyk2, and of the double deletion mutant DELTApyk1DELTApyk2. Deletion of pyk1 results in marginal Pyk activity that is below the detection limit, complementation of the DELTApyk1DELTApyk2 strain with the pyk2 gene | Corynebacterium glutamicum |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| ATP | allosteric regulation | Corynebacterium glutamicum | |
| additional information | glucose-6-phosphate (G6P) has no significant effect on the S0.5 and kcat of Pyk2 | Corynebacterium glutamicum |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | isozyme Pyk2 has a relatively lower affinity for ADP compared to isozyme Pyk1 | Corynebacterium glutamicum | |
| 0.07 | - |
ADP | pH 7.0, 35°C | Corynebacterium glutamicum | |
| 0.33 | - |
ADP | pH 7.0, 35°C | Corynebacterium glutamicum | |
| 4.58 | - |
phosphoenolpyruvate | pH 7.0, 35°C | Corynebacterium glutamicum | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | activates | Corynebacterium glutamicum | |
| K+ | Pyk2 is co-dependent on Mn2+ and K+ | Corynebacterium glutamicum | |
| Li+ | can substitute for K+ by 38% | Corynebacterium glutamicum | |
| Mn2+ | activates | Corynebacterium glutamicum | |
| Mn2+ | Pyk2 is co-dependent on Mn2+ and K+ | Corynebacterium glutamicum | |
| additional information | isozyme Pyk2 exhibits an absolute dependence on Mn2+ together with a monovalent cation. The maximum activity of Pyk2 is detected in the presence of 5 mM Mn2+ and 100 mM K+. When NH4 +, Na+, or Li+ is substituted for K+, the relative activities of Pyk2 are maintained at 73, 51, and 38%, respectively | Corynebacterium glutamicum | |
| additional information | Pyk1 exhibits high catalytic activity using Mn2+ or Co2+ as a cation, Pyk1 only uses Mn2+ or Co2+ as cations | Corynebacterium glutamicum | |
| Na+ | can substitute for K+ by 51% | Corynebacterium glutamicum | |
| NH4+ | can substitute for K+ by 73% | Corynebacterium glutamicum | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 430881 | - |
gel filtration | Corynebacterium glutamicum |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ADP + phosphoenolpyruvate | Corynebacterium glutamicum | - |
ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | Corynebacterium glutamicum LMG 3730 | - |
ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | Corynebacterium glutamicum BCRC 11384 | - |
ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | Corynebacterium glutamicum ATCC 13032 | - |
ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | Corynebacterium glutamicum JCM 1318 | - |
ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | Corynebacterium glutamicum NCIMB 10025 | - |
ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | Corynebacterium glutamicum DSM 20300 | - |
ATP + pyruvate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Corynebacterium glutamicum | Q46078 | - |
- |
| Corynebacterium glutamicum | Q8NLN1 | - |
- |
| Corynebacterium glutamicum ATCC 13032 | Q46078 | - |
- |
| Corynebacterium glutamicum ATCC 13032 | Q8NLN1 | - |
- |
| Corynebacterium glutamicum BCRC 11384 | Q46078 | - |
- |
| Corynebacterium glutamicum BCRC 11384 | Q8NLN1 | - |
- |
| Corynebacterium glutamicum DSM 20300 | Q46078 | - |
- |
| Corynebacterium glutamicum DSM 20300 | Q8NLN1 | - |
- |
| Corynebacterium glutamicum JCM 1318 | Q46078 | - |
- |
| Corynebacterium glutamicum JCM 1318 | Q8NLN1 | - |
- |
| Corynebacterium glutamicum LMG 3730 | Q46078 | - |
- |
| Corynebacterium glutamicum LMG 3730 | Q8NLN1 | - |
- |
| Corynebacterium glutamicum NCIMB 10025 | Q46078 | - |
- |
| Corynebacterium glutamicum NCIMB 10025 | Q8NLN1 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant N-terminally His6-tagged enzyme 12.81fold from Escherichia coli strain BL21(DE3) by nickel affinity and ion exchange chromatography | Corynebacterium glutamicum |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 1.64 | - |
purified recombinant enzyme, pH 7.0, 35°C | Corynebacterium glutamicum |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ADP + phosphoenolpyruvate | - |
Corynebacterium glutamicum | ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | - |
Corynebacterium glutamicum LMG 3730 | ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | - |
Corynebacterium glutamicum BCRC 11384 | ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | - |
Corynebacterium glutamicum ATCC 13032 | ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | - |
Corynebacterium glutamicum JCM 1318 | ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | - |
Corynebacterium glutamicum NCIMB 10025 | ATP + pyruvate | - |
? | |
| ADP + phosphoenolpyruvate | - |
Corynebacterium glutamicum DSM 20300 | ATP + pyruvate | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homohexamer | 6 * 67572, sequence calculation | Corynebacterium glutamicum |
| More | a long extra N-terminal sequence of 120 amino acids makes Pyk2 (67.6 kDa) larger than other typical bacterial Pyks (51 kDa), most other Pyk2s are homotetramers, enzyme structure comparisons | Corynebacterium glutamicum |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| NCgl2008 | locus name | Corynebacterium glutamicum |
| NCgl2809 | locus name | Corynebacterium glutamicum |
| PYK | - |
Corynebacterium glutamicum |
| Pyk1 | - |
Corynebacterium glutamicum |
| Pyk2 | - |
Corynebacterium glutamicum |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 35 | - |
- |
Corynebacterium glutamicum |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 20 | 50 | activity range, profile overview | Corynebacterium glutamicum |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 4.36 | - |
ADP | pH 7.0, 35°C | Corynebacterium glutamicum | |
| 5.58 | - |
phosphoenolpyruvate | pH 7.0, 35°C | Corynebacterium glutamicum | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
- |
Corynebacterium glutamicum |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 9 | over 80% of maximal activity within this range, profile overview | Corynebacterium glutamicum |
Expression
| Organism | Comment | Expression |
|---|---|---|
| Corynebacterium glutamicum | the mRNA level of pyk2 in the wild-type strain under oxygen deprivation is 14.24fold higher than that under aerobic conditions. Expression of pyk2 is induced under oxygen deprivation | up |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | although Corynebacterium glutamicum is assumed to possess only one Pyk (pyk1, NCgl2008), NCgl2809 is annotated as a pyruvate kinase with an unknown role. NCgl2809 is identified as encoding pyruvate kinase (pyk2) in Corynebacterium glutamicum | Corynebacterium glutamicum |
| malfunction | complementation of the DELTApyk1DELTApyk2 strain with the pyk2 gene restores its growth on D-ribose, which demonstrates that Pyk2 can substitute for Pyk1 in vivo. Under oxygen deprivation, pyk1 or pyk2 deficiency decreases the generation of lactic acid, and the overexpression of either pyk1 or pyk2 increases the production of lactic acid as the activity of Pyk increases. Fed-batch fermentation of the pyk2-overexpressing WTDELTApyk1 strain produces 60.27 g/l of lactic acid, which is a 47% increase compared to the parent strain under oxygen deprivation | Corynebacterium glutamicum |
| malfunction | deletion of pyk1 results in marginal Pyk activity that is below the detection limit. Complementation of the DELTApyk1DELTApyk2 strain with the pyk2 gene restores its growth on D-ribose, which demonstrates that Pyk2 can substitute for Pyk1 in vivo. Under oxygen deprivation, pyk1 or pyk2 deficiency decreases the generation of lactic acid, and the overexpression of either pyk1 or pyk2 increases the production of lactic acid as the activity of Pyk increases | Corynebacterium glutamicum |
| physiological function | pyruvate kinase (Pyk) catalyzes the generation of pyruvate and ATP in glycolysis and functions as a key switch in the regulation of carbon flux distribution. Both the substrates and products of Pyk are involved in the tricarboxylic acid cycle, anaplerosis and energy anabolism, which places Pyk at a primary metabolic intersection | Corynebacterium glutamicum |
| physiological function | pyruvate kinase (Pyk) catalyzes the generation of pyruvate and ATP in glycolysis and functions as a key switch in the regulation of carbon flux distribution. Both the substrates and products of Pyk are involved in the tricarboxylic acid cycle, anaplerosis and energy anabolism, which places Pyk at a primary metabolic intersection. Pyk2 functions as a pyruvate kinase and contributes to the increased level of Pyk activity under oxygen deprivation. The catalytic activity of Pyk2 is allosterically regulated by fructose 1,6-bisphosphate activation and ATP inhibition | Corynebacterium glutamicum |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.22 | - |
phosphoenolpyruvate | pH 7.0, 35°C | Corynebacterium glutamicum | |
| 13.2 | - |
ADP | pH 7.0, 35°C | Corynebacterium glutamicum | |
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Release 2023.1 (January 2023)
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