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Literature summary for 2.7.1.40 extracted from

  • Sakai, H.
    Mutagenesis of the active site lysine 221 of the pyruvate kinase from Bacillus stearothermophilus (2005), J. Biochem., 137, 141-145.
    View publication on PubMed
Search for more literature for 2.7.1.40

Activating Compound

Activating Compound Comment Organism Structure
D-ribose 5-phosphate 0.1 mM, allosteric activator Geobacillus stearothermophilus

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli XL-1 Blue cells Geobacillus stearothermophilus

Protein Variants

Protein Variants Comment Organism
C9S/C268S 75% activity of the wild-type enzyme Geobacillus stearothermophilus
C9S/C268S/K221C 10000 times less active than the wild type enzyme Geobacillus stearothermophilus
K221C 10000 to 100000 times less active than the wild-type enzyme Geobacillus stearothermophilus
K221D 10000 to 100000 times less active than the wild-type enzyme Geobacillus stearothermophilus
K221L 10000 to 100000 times less active than the wild-type enzyme Geobacillus stearothermophilus
K221R 10000 to 100000 times less active than the wild-type enzyme Geobacillus stearothermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.113
-
 phosphoenolpyruvate mutant enzyme C9S/C268S, in the presence of 0.1 mM ribose 5'-phosphate, in 50 mM imidazole-HCl buffer, pH 7.2, 50 mM KCl, 7 mM MgCl2, 0.12 mM NADH, 0.02 mg/ml lactate dehydrogenase, 2 mM PEP, and 4 mM ADP, at 30°C Geobacillus stearothermophilus
0.186
-
 phosphoenolpyruvate wild type enzyme, in the presence of 0.1 mM ribose 5'-phosphate, in 50 mM imidazole-HCl buffer, pH 7.2, 50 mM KCl, 7 mM MgCl2, 0.12 mM NADH, 0.02 mg/ml lactate dehydrogenase, 2 mM PEP, and 4 mM ADP, at 30°C Geobacillus stearothermophilus

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Butyl-Toyopearl 650S column chromatography and MonoQ HR10/10 column chromatography Geobacillus stearothermophilus

Renatured (Commentary)

Renatured (Comment) Organism
dialysis against 0.2 M Tris-HCl, pH 8.6, containing 1 mM EDTA and 0.05 mM dithiothreitol, after denaturation with 4 M guanidine hydrochloride Geobacillus stearothermophilus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 555 units/mg, wild type enzyme, at 30°C Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP + phosphoenolpyruvate
-
 Geobacillus stearothermophilus ATP + pyruvate
-
 ?

Synonyms

Synonyms Comment Organism
phosphoenolpyruvate kinase
-
 Geobacillus stearothermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1736
-
 phosphoenolpyruvate mutant enzyme C9S/C268S, in the presence of 0.1 mM ribose 5'-phosphate, in 50 mM imidazole-HCl buffer, pH 7.2, 50 mM KCl, 7 mM MgCl2, 0.12 mM NADH, 0.02 mg/ml lactate dehydrogenase, 2 mM PEP, and 4 mM ADP, at 30°C Geobacillus stearothermophilus
3204
-
 phosphoenolpyruvate wild type enzyme, in the presence of 0.1 mM ribose 5'-phosphate, in 50 mM imidazole-HCl buffer, pH 7.2, 50 mM KCl, 7 mM MgCl2, 0.12 mM NADH, 0.02 mg/ml lactate dehydrogenase, 2 mM PEP, and 4 mM ADP, at 30°C Geobacillus stearothermophilus

Cofactor

Cofactor Comment Organism Structure
ADP
-
 Geobacillus stearothermophilus