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Literature summary for 1.1.1.28 extracted from
- The D-lactate dehydrogenase from Sporolactobacillus inulinus also possessing reversible deamination activity (2015), PLoS ONE, 10, e0139066.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene ldhD, phylogenetic analysis, recombinant expression of wwild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Sporolactobacillus inulinus |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| the complete tertiary structure of DLDH744 in complex with NAD+ is determined, PDB ID 4XKJ, determined by molecular replacement method and refined at 3.15 A-resolution | Sporolactobacillus inulinus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| E263A | site-directed mutagenesis, the mutant shows highly reduced D-lactate dehydrogenase activity compared to the wild-type enzyme | Sporolactobacillus inulinus |
| F298A | site-directed mutagenesis, almost inactive D-lactate dehydrogenase mutant | Sporolactobacillus inulinus |
| G79A | site-directed mutagenesis, almost inactive D-lactate dehydrogenase mutant | Sporolactobacillus inulinus |
| H295A | site-directed mutagenesis, inactive D-lactate dehydrogenase mutant | Sporolactobacillus inulinus |
| M307A | site-directed mutagenesisthe mutant shows highly reduced D-lactate dehydrogenase activity compared to the wild-type enzyme | Sporolactobacillus inulinus |
| additional information | glutamate dehydrogenase activities of enzyme mutants, overview | Sporolactobacillus inulinus |
| R234A | site-directed mutagenesis, inactive D-lactate dehydrogenase mutant | Sporolactobacillus inulinus |
| Y101A | site-directed mutagenesis, the mutant shows 50% reduced D-lactate dehydrogenase activity compared to the wild-type enzyme | Sporolactobacillus inulinus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 3.32 | - |
phenylpyruvate | recombinant enzyme, pH and temperature not specified in the publication | Sporolactobacillus inulinus |  |
| 3.4 | - |
pyruvate | recombinant enzyme, pH and temperature not specified in the publication | Sporolactobacillus inulinus | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Sporolactobacillus inulinus | A0A0M3KL04 | - |
- |
| Sporolactobacillus inulinus CASD | A0A0M3KL04 | - |
- |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| (R)-lactate + NAD+ = pyruvate + NADH + H+ | catalytic mechanism | Sporolactobacillus inulinus |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
purified recombinant enzyme, pH and temperature not specified in the publication | Sporolactobacillus inulinus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (R)-lactate + NAD(P)+ | the enzyme is confirmed to be a typical D-lactate dehydrogenase based on the activity detected with substrate pyruvate and coenzyme NAD(P)H | Sporolactobacillus inulinus | pyruvate + NAD(P)H + H+ | - |
r | |
| (R)-lactate + NAD(P)+ | the enzyme is confirmed to be a typical D-lactate dehydrogenase based on the activity detected with substrate pyruvate and coenzyme NAD(P)H | Sporolactobacillus inulinus CASD | pyruvate + NAD(P)H + H+ | - |
r | |
| additional information | the bifunctional enzyme from Sporolactobacillus inulinus has both the D-LDH and NAD(P)+/NAD(P)H-utilizing glutamate dehydrogenase (GDH, EC 1.4.1.3) activities, the latter with reversible deamination. Identification of key residues from the crystal structure analysis and site-directed mutagenesis, enzyme reidues Arg234 and Gly79 residues play a significant role in both D-LDH and GDH activities. Residues His295 and Phe298 in DLDH744 are identified to be key residues for lactate dehydrogenase (LDH) activity only, whereas Tyr101 is a unique residue that is critical for GDH activity. DLDH744 is a strict D-2-hydroxyacid dehydrogenase, no formation of L-isomers | Sporolactobacillus inulinus | ? | - |
? | |
| additional information | the bifunctional enzyme from Sporolactobacillus inulinus has both the D-LDH and NAD(P)+/NAD(P)H-utilizing glutamate dehydrogenase (GDH, EC 1.4.1.3) activities, the latter with reversible deamination. Identification of key residues from the crystal structure analysis and site-directed mutagenesis, enzyme reidues Arg234 and Gly79 residues play a significant role in both D-LDH and GDH activities. Residues His295 and Phe298 in DLDH744 are identified to be key residues for lactate dehydrogenase (LDH) activity only, whereas Tyr101 is a unique residue that is critical for GDH activity. DLDH744 is a strict D-2-hydroxyacid dehydrogenase, no formation of L-isomers | Sporolactobacillus inulinus CASD | ? | - |
? | |
| phenylpyruvate + NADH + H+ | - |
Sporolactobacillus inulinus | D-phenyllactate + NAD+ | - |
r | |
| phenylpyruvate + NADH + H+ | - |
Sporolactobacillus inulinus CASD | D-phenyllactate + NAD+ | - |
r | |
| pyruvate + NAD(P)H + H+ | - |
Sporolactobacillus inulinus | (R)-lactate + NAD(P)+ | - |
r | |
| pyruvate + NAD(P)H + H+ | - |
Sporolactobacillus inulinus CASD | (R)-lactate + NAD(P)+ | - |
r | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homodimer | two identical monomers of DLDH744 are associated at the cofactor binding domain to form the homodimer in one asymmetric unit. The dimer interface is stabilized mainly by hydrogen-bonding interactions between residues from alpha-helices, alpha5(Ser102-Arg119), alpa6(Leu121-Glu130), alpha12(Glu279-Arg285) and connecting loops, structure analysis, overview. His295 is supposed to function as the internal acid-base catalyst in dehydrogenase activity. Glu263 stabilizes the protonated form of His295, and Arg234 forms a hydrogen bond with the nicotinamide ribose ring | Sporolactobacillus inulinus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| D-LDH | - |
Sporolactobacillus inulinus |
| DLDH744 | - |
Sporolactobacillus inulinus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 512.2 | - |
pyruvate | recombinant enzyme, pH and temperature not specified in the publication | Sporolactobacillus inulinus | |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NAD+ | - |
Sporolactobacillus inulinus | |
| NADH | - |
Sporolactobacillus inulinus | |
| NADP+ | - |
Sporolactobacillus inulinus | |
| NADPH | - |
Sporolactobacillus inulinus | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to the D-isomer-specific 2-hydroxyacid dehydrogenase family | Sporolactobacillus inulinus |
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