Activating Compound | Comment | Organism | Structure |
---|---|---|---|
D-fructose 1,6-bisphosphate | activates the pyruvate reduction | Geobacillus stearothermophilus |
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 (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain JM109 | Geobacillus stearothermophilus |
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 | 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 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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-lactate + NAD+ | Geobacillus stearothermophilus | - |
pyruvate + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus stearothermophilus | - |
- |
- |
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 | 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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-lactate + NAD+ | - |
Geobacillus stearothermophilus | pyruvate + NADH + H+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
LDH | - |
Geobacillus stearothermophilus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at, reduction reaction | Geobacillus stearothermophilus |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at, reduction reaction | Geobacillus stearothermophilus |
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 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 |