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Literature summary for 1.1.1.27 extracted from

  • Binay, B.; Karagueler, N.G.
    Attempting to remove the substrate inhibition of L-lactate dehydrogenase from Bacillus stearothermophilus by site-directed mutagenesis (2007), Appl. Biochem. Biotechnol., 141, 265-272.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
D-fructose 1,6-bisphosphate activates the pyruvate reduction Geobacillus stearothermophilus

Application

Application Comment Organism
synthesis the enzyme has a commercial significance, as it can be used to produce chiral building blocks for the synthesis of key pharmaceuticals and agrochemicals, optimization of enzyme reaction by engineering to eliminate the substrate inhibition Geobacillus stearothermophilus

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain JM109 Geobacillus stearothermophilus

Protein Variants

Protein Variants Comment Organism
D38E site-directed mutagenesis, the mutant shows a twofold reduced substrate inhibition by pyruvate compared to the wild-type enzyme Geobacillus stearothermophilus
D38R site-directed mutagenesis, the mutant shows a threefold reduced substrate inhibition by pyruvate compared to the wild-type enzyme Geobacillus stearothermophilus

Inhibitors

Inhibitors Comment Organism Structure
NAD+ substrate inhibition due to an abortive NAD+-pyruvate complex reducing the steady state concentration of functional LDH Geobacillus stearothermophilus
pyruvate substrate inhibition due to an abortive NAD+-pyruvate complex reducing the steady state concentration of functional LDH Geobacillus stearothermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.76
-
pyruvate pH 6.0, 25°C, recombinant wild-type enzyme in presence of fructose 1,6-bisphosphate Geobacillus stearothermophilus
3.7
-
pyruvate pH 6.0, 25°C, recombinant mutant D38R in presence of fructose 1,6-bisphosphate Geobacillus stearothermophilus
19.3
-
pyruvate pH 6.0, 25°C, recombinant mutant D38R in absence of fructose 1,6-bisphosphate Geobacillus stearothermophilus
32
-
pyruvate pH 6.0, 25°C, recombinant wild-type enzyme in absence of fructose 1,6-bisphosphate Geobacillus stearothermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-lactate + NAD+ Geobacillus stearothermophilus
-
pyruvate + NADH + H+
-
r

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain JM109 by ion-exchange chromatography and affinity chromatography, respectively Geobacillus stearothermophilus

Reaction

Reaction Comment Organism Reaction ID
(S)-lactate + NAD+ = pyruvate + NADH + H+ the NAD (H)-dependent L-LDH catalyzes the reduction of pyruvate by an ordered catalytic mechanism, catalytic cycle, overview Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-lactate + NAD+
-
Geobacillus stearothermophilus pyruvate + NADH + H+
-
r

Synonyms

Synonyms Comment Organism
LDH
-
Geobacillus stearothermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at, reduction reaction Geobacillus stearothermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
21.1
-
pyruvate pH 6.0, 25°C, recombinant wild-type enzyme in presence of fructose 1,6-bisphosphate Geobacillus stearothermophilus
178.4
-
pyruvate pH 6.0, 25°C, recombinant mutant D38R in presence of fructose 1,6-bisphosphate Geobacillus stearothermophilus
194.9
-
pyruvate pH 6.0, 25°C, recombinant mutant D38R in absence of fructose 1,6-bisphosphate Geobacillus stearothermophilus
327.2
-
pyruvate pH 6.0, 25°C, recombinant wild-type enzyme in absence of fructose 1,6-bisphosphate Geobacillus stearothermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
assay at, reduction reaction Geobacillus stearothermophilus

Cofactor

Cofactor Comment Organism Structure
NAD+ the coenzyme is in an open conformation and the adenine ribose ring of it is surrounded by Asp 38, Val 39, and Gly 99. Asp 38 and Gly 99 give some specificity to the adenine orientation, overview Geobacillus stearothermophilus
NADH the coenzyme is in an open conformation and the adenine ribose ring of it is surrounded by Asp 38, Val 39, and Gly 99. Asp 38 and Gly 99 give some specificity to the adenine orientation, Asp 38 is an important residue in stabilizing NADH binding, overview Geobacillus stearothermophilus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
10.9
-
pyruvate pH 6.0, 25°C, recombinant wild-type enzyme in presence of fructose 1,6-bisphosphate Geobacillus stearothermophilus
31.1
-
pyruvate pH 6.0, 25°C, recombinant mutant D38R in presence of fructose 1,6-bisphosphate Geobacillus stearothermophilus