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Literature summary extracted from

  • Zhu, L.; Xu, X.; Wang, L.; Dong, H.; Yu, B.
    The D-lactate dehydrogenase from Sporolactobacillus inulinus also possessing reversible deamination activity (2015), PLoS ONE, 10, e0139066.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.28 gene ldhD, phylogenetic analysis, recombinant expression of wwild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Sporolactobacillus inulinus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.28 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

EC Number Protein Variants Comment Organism
1.1.1.28 E263A site-directed mutagenesis, the mutant shows highly reduced D-lactate dehydrogenase activity compared to the wild-type enzyme Sporolactobacillus inulinus
1.1.1.28 F298A site-directed mutagenesis, almost inactive D-lactate dehydrogenase mutant Sporolactobacillus inulinus
1.1.1.28 G79A site-directed mutagenesis, almost inactive D-lactate dehydrogenase mutant Sporolactobacillus inulinus
1.1.1.28 H295A site-directed mutagenesis, inactive D-lactate dehydrogenase mutant Sporolactobacillus inulinus
1.1.1.28 M307A site-directed mutagenesisthe mutant shows highly reduced D-lactate dehydrogenase activity compared to the wild-type enzyme Sporolactobacillus inulinus
1.1.1.28 additional information glutamate dehydrogenase activities of enzyme mutants, overview Sporolactobacillus inulinus
1.1.1.28 R234A site-directed mutagenesis, inactive D-lactate dehydrogenase mutant Sporolactobacillus inulinus
1.1.1.28 Y101A site-directed mutagenesis, the mutant shows 50% reduced D-lactate dehydrogenase activity compared to the wild-type enzyme Sporolactobacillus inulinus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.28 3.32
-
 phenylpyruvate recombinant enzyme, pH and temperature not specified in the publication Sporolactobacillus inulinus
1.1.1.28 3.4
-
 pyruvate recombinant enzyme, pH and temperature not specified in the publication Sporolactobacillus inulinus

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.28 Sporolactobacillus inulinus A0A0M3KL04
-
-
1.1.1.28 Sporolactobacillus inulinus CASD A0A0M3KL04
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.28 (R)-lactate + NAD+ = pyruvate + NADH + H+ catalytic mechanism Sporolactobacillus inulinus
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.28 7.5
-
 purified recombinant enzyme, pH and temperature not specified in the publication Sporolactobacillus inulinus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.28 (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
1.1.1.28 (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
1.1.1.28 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 ?
-
 ?
1.1.1.28 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 ?
-
 ?
1.1.1.28 phenylpyruvate + NADH + H+
-
 Sporolactobacillus inulinus D-phenyllactate + NAD+
-
 r
1.1.1.28 phenylpyruvate + NADH + H+
-
 Sporolactobacillus inulinus CASD D-phenyllactate + NAD+
-
 r
1.1.1.28 pyruvate + NAD(P)H + H+
-
 Sporolactobacillus inulinus (R)-lactate + NAD(P)+
-
 r
1.1.1.28 pyruvate + NAD(P)H + H+
-
 Sporolactobacillus inulinus CASD (R)-lactate + NAD(P)+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.28 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

EC Number Synonyms Comment Organism
1.1.1.28 D-LDH
-
 Sporolactobacillus inulinus
1.1.1.28 DLDH744
-
 Sporolactobacillus inulinus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.28 512.2
-
 pyruvate recombinant enzyme, pH and temperature not specified in the publication Sporolactobacillus inulinus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.28 NAD+
-
 Sporolactobacillus inulinus
1.1.1.28 NADH
-
 Sporolactobacillus inulinus
1.1.1.28 NADP+
-
 Sporolactobacillus inulinus
1.1.1.28 NADPH
-
 Sporolactobacillus inulinus

General Information

EC Number General Information Comment Organism
1.1.1.28 evolution the enzyme belongs to the D-isomer-specific 2-hydroxyacid dehydrogenase family Sporolactobacillus inulinus