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Literature summary extracted from

  • Puri, P.; Goel, A.; Bochynska, A.; Poolman, B.
    Regulation of acetate kinase isozymes and its importance for mixed-acid fermentation in Lactococcus lactis (2014), J. Bacteriol., 196, 1386-1393.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.2.1 gene acka1, recombinant expression of His-tagged isozyme AckA1 in Escherichia coli strain MC1061 Lactococcus lactis
2.7.2.1 gene acka2, recombinant expression of His-tagged isozyme AckA2 in Escherichia coli strain MC1061 Lactococcus lactis

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.2.1 ADP competitive inhibition; competitive inhibition Lactococcus lactis
2.7.2.1 D-fructose-1,6-bisphosphate inhibits the activities of isozymes AckA1 and AckA2; inhibits the activities of isozymes AckA1 and AckA2 Lactococcus lactis
2.7.2.1 glyceraldehyde-3-phosphate inhibits the activities of isozyme AckA1, but very poorly of isozyme AckA2; inhibits the activities of isozyme AckA1, but very poorly of isozyme AckA2 Lactococcus lactis
2.7.2.1 phospho-enol-pyruvate PEP, a downstream intermediate of glycolysis, completely inhibits the activity of both enzymes at concentrations above 30 mM; PEP, a downstream intermediate of glycolysis, completely inhibits the activity of both enzymes at concentrations above 30 mM Lactococcus lactis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.2.1 additional information
-
 additional information the turnover number of AckA1 is about an order of magnitude higher than that of AckA2 for the reaction in either direction. The Km values for acetyl phosphate, ATP, and ADP are similar for both isozymes. AckA2 has a higher affinity for acetate than does AckA1. Michaelis-Menten kinetics Lactococcus lactis
2.7.2.1 0.07
-
 ATP pH 7.5, 30°C, recombinant His-tagged isozyme AckA1 Lactococcus lactis
2.7.2.1 0.07
-
 ATP pH 7.5, 30°C, recombinant His-tagged isozyme AckA2 Lactococcus lactis
2.7.2.1 0.47
-
 ADP pH 7.5, 30°C, recombinant His-tagged isozyme AckA1 Lactococcus lactis
2.7.2.1 0.54
-
 acetyl phosphate pH 7.5, 30°C, recombinant His-tagged isozyme AckA1 Lactococcus lactis
2.7.2.1 0.55
-
 acetyl phosphate pH 7.5, 30°C, recombinant His-tagged isozyme AckA2 Lactococcus lactis
2.7.2.1 0.74
-
 ADP pH 7.5, 30°C, recombinant His-tagged isozyme AckA2 Lactococcus lactis
2.7.2.1 4.9
-
 acetate pH 7.5, 30°C, recombinant His-tagged isozyme AckA2 Lactococcus lactis
2.7.2.1 20.54
-
 acetate pH 7.5, 30°C, recombinant His-tagged isozyme AckA1 Lactococcus lactis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.2.1 K+ the enzyme requires monovalent K+ and divalent Mn2+or Mg2+ cations Lactococcus lactis
2.7.2.1 Mg2+ the enzyme requires monovalent K+ and divalent Mn2+or Mg2+ cations Lactococcus lactis
2.7.2.1 Mn2+ the enzyme requires monovalent K+ and divalent Mn2+or Mg2+ cations Lactococcus lactis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.2.1 ATP + acetate Lactococcus lactis
-
 ADP + acetyl phosphate
-
 r
2.7.2.1 ATP + acetate Lactococcus lactis MG1363
-
 ADP + acetyl phosphate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.7.2.1 Lactococcus lactis A2RNG4 gene acka2; isozyme AckA2 encoded by gene acka2
-
2.7.2.1 Lactococcus lactis A2RNG5 gene acka1; isozyme AckA1 encoded by gene acka1
-
2.7.2.1 Lactococcus lactis MG1363 A2RNG4 gene acka2; isozyme AckA2 encoded by gene acka2
-
2.7.2.1 Lactococcus lactis MG1363 A2RNG5 gene acka1; isozyme AckA1 encoded by gene acka1
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.2.1 recombinant His-tagged isozyme AckA1 from Escherichia coli strain MC1061 by nickel affinity chromatography Lactococcus lactis
2.7.2.1 recombinant His-tagged isozyme AckA2 from Escherichia coli strain MC1061 by nickel affinity chromatography Lactococcus lactis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.2.1 ATP + acetate
-
 Lactococcus lactis ADP + acetyl phosphate
-
 r
2.7.2.1 ATP + acetate
-
 Lactococcus lactis MG1363 ADP + acetyl phosphate
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.7.2.1 ACK
-
 Lactococcus lactis
2.7.2.1 AckA1
-
 Lactococcus lactis
2.7.2.1 AckA2
-
 Lactococcus lactis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.2.1 30
-
 assay at Lactococcus lactis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.2.1 80
-
 acetate pH 7.5, 30°C, recombinant His-tagged isozyme AckA2 Lactococcus lactis
2.7.2.1 93
-
 acetyl phosphate pH 7.5, 30°C, recombinant His-tagged isozyme AckA2 Lactococcus lactis
2.7.2.1 761
-
 acetate pH 7.5, 30°C, recombinant His-tagged isozyme AckA1 Lactococcus lactis
2.7.2.1 1105
-
 acetyl phosphate pH 7.5, 30°C, recombinant His-tagged isozyme AckA1 Lactococcus lactis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.2.1 7.5
-
 assay at Lactococcus lactis

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.2.1 ADP
-
 Lactococcus lactis
2.7.2.1 ATP
-
 Lactococcus lactis

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.7.2.1 2.4
-
 ADP pH 7.5, 30°C, recombinant His-tagged isozyme AckA1 Lactococcus lactis
2.7.2.1 3.9
-
 ADP pH 7.5, 30°C, recombinant His-tagged isozyme AckA2 Lactococcus lactis
2.7.2.1 4
-
 glyceraldehyde-3-phosphate pH 7.5, 30°C, recombinant His-tagged isozyme AckA1 Lactococcus lactis
2.7.2.1 15
-
 phospho-enol-pyruvate pH 7.5, 30°C, recombinant His-tagged isozyme AckA1 Lactococcus lactis
2.7.2.1 17
-
 D-fructose-1,6-bisphosphate pH 7.5, 30°C, recombinant His-tagged isozyme AckA1 Lactococcus lactis
2.7.2.1 18
-
 phospho-enol-pyruvate pH 7.5, 30°C, recombinant His-tagged isozyme AckA2 Lactococcus lactis
2.7.2.1 43
-
 D-fructose-1,6-bisphosphate pH 7.5, 30°C, recombinant His-tagged isozyme AckA2 Lactococcus lactis

General Information

EC Number General Information Comment Organism
2.7.2.1 physiological function acetate kinase converts acetyl phosphate to acetate along with the generation of ATP in the pathway for mixed-acid fermentation in Lactococcus lactis. The reverse reaction yields acetyl phosphate for assimilation purposes. AckA1 is an order of magnitude more active than AckA2 in both reaction directions, but AckA2 has the highest affinity for acetate. Allosteric regulation of isozymes AckA1 and AckA2 and the pool sizes of the glycolytic intermediates are consistent with a switch from homolactic to mixed-acid fermentation upon slowing of the growth rate Lactococcus lactis
2.7.2.1 physiological function acetate kinase converts acetyl phosphate to acetate along with the generation of ATP in the pathway for mixed-acid fermentation in Lactococcus lactis. The reverse reaction yields acetyl phosphate for assimilation purposes. AckA1 is an order of magnitude more active than AckA2 in both reaction directions, but AckA2 has the highest affinity for acetate. Isozyme AckA2 has an important role under acetate-limiting conditions despite the lower activity. Allosteric regulation of isozymes AckA1 and AckA2 and the pool sizes of the glycolytic intermediates are consistent with a switch from homolactic to mixed-acid fermentation upon slowing of the growth rate Lactococcus lactis