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Literature summary extracted from

  • Domenech, J.; Ferrer, J.
    A new D-2-hydroxyacid dehydrogenase with dual coenzyme-specificity from Haloferax mediterranei, sequence analysis and heterologous overexpression (2006), Biochim. Biophys. Acta, 1760, 1667-1674.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.272 gene ddh, DNA and amino acid sequence determination and analysis, sequence comparison, subcloning and expression in Escherichia coli strains XL1 Blue and BL21(DE3) in inclusion bodies Haloferax mediterranei

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.272 0.031
-
 NADPH pH 8.5, 40°C, recombinant enzyme, substrate 2-oxoisocaproate Haloferax mediterranei
1.1.1.272 0.046
-
 NADPH pH 8.5, 40°C, recombinant enzyme, substrate 2-oxoisoleucine Haloferax mediterranei
1.1.1.272 0.054
-
 NADPH pH 8.5, 40°C, recombinant enzyme, substrate 2-oxobutyrate Haloferax mediterranei
1.1.1.272 0.233
-
 NADH pH 8.5, 40°C, recombinant enzyme, substrate 2-oxoisocaproate Haloferax mediterranei
1.1.1.272 0.33
-
 NADH pH 8.5, 40°C, recombinant enzyme, substrate 2-oxoisoleucine Haloferax mediterranei
1.1.1.272 0.5
-
 NADH pH 8.5, 40°C, recombinant enzyme, substrate 2-oxobutyrate Haloferax mediterranei
1.1.1.272 1.31
-
 2-oxoisoleucine pH 8.5, 40°C, recombinant enzyme, cofactor NADPH Haloferax mediterranei
1.1.1.272 3.77
-
 2-oxoisocaproate pH 8.5, 40°C, recombinant enzyme, cofactor NADPH Haloferax mediterranei
1.1.1.272 11.9
-
 2-oxoisoleucine pH 8.5, 40°C, recombinant enzyme, cofactor NADH Haloferax mediterranei
1.1.1.272 13.45
-
 2-oxobutyrate pH 8.5, 40°C, recombinant enzyme, cofactor NADPH Haloferax mediterranei
1.1.1.272 13.9
-
 2-oxoisocaproate pH 8.5, 40°C, recombinant enzyme, cofactor NADH Haloferax mediterranei
1.1.1.272 21.96
-
 pyruvate pH 5.0, 40°C, recombinant enzyme, cofactor NADPH Haloferax mediterranei
1.1.1.272 106
-
 2-oxobutyrate pH 8.5, 40°C, recombinant enzyme, cofactor NADH Haloferax mediterranei

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.272 29500
-
 4 * 33336, sequence calculation, 4 * 29500, recombinant enzyme, CTAB-PAGE, 4 * 47000, SDS-PAGE Haloferax mediterranei
1.1.1.272 33336
-
 4 * 33336, sequence calculation, 4 * 29500, recombinant enzyme, CTAB-PAGE, 4 * 47000, SDS-PAGE Haloferax mediterranei
1.1.1.272 47000
-
 4 * 33336, sequence calculation, 4 * 29500, recombinant enzyme, CTAB-PAGE, 4 * 47000, SDS-PAGE Haloferax mediterranei
1.1.1.272 111000
-
 recombinant enzyme, gel filtration Haloferax mediterranei

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.272 Haloferax mediterranei Q2VEQ7 gene ddh; strain R4, gene ddh
-
1.1.1.272 Haloferax mediterranei R-4 / ATCC 33500 Q2VEQ7 gene ddh; strain R4, gene ddh
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.272 recombinant enzyme partially from Escherichia coli strain BL21(DE3) inclusion bodies by solubilization with 8 M urea in DTT-containing buffer and refolding by rapid 10fold dilution Haloferax mediterranei

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.272 an (R)-2-hydroxycarboxylate + NADP+ = a 2-oxocarboxylate + NADPH + H+ the catalytic triad is probably formed by Arg226, Glu255, and His274 Haloferax mediterranei
a

Renatured (Commentary)

EC Number Renatured (Comment) Organism
1.1.1.272 refolding of recombinant enzyme from Escherichia coli strain BL21(DE3) inclusion bodies, solubilized by 8 M urea in DTT-containing buffer, by rapid 10fold dilution, optimally in presence of 4 M NaCl at pH 8.0 and 37°C Haloferax mediterranei

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.272 additional information
-
 substrate and cofactor specificity, overview Haloferax mediterranei

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.272 2-oxo-4-methylpentanoate + NAD(P)H
-
 Haloferax mediterranei (2R)-2-hydroxy-4-methylpentanoate + NAD(P)+
-
 ir
1.1.1.272 2-oxo-4-methylpentanoate + NAD(P)H
-
 Haloferax mediterranei R-4 / ATCC 33500 (2R)-2-hydroxy-4-methylpentanoate + NAD(P)+
-
 ir
1.1.1.272 2-oxobutyrate + NAD(P)H
-
 Haloferax mediterranei (2R)-2-hydroxybutyrate + NAD(P)+
-
 ir
1.1.1.272 2-oxobutyrate + NAD(P)H
-
 Haloferax mediterranei R-4 / ATCC 33500 (2R)-2-hydroxybutyrate + NAD(P)+
-
 ir
1.1.1.272 2-oxobutyrate + NADH + H+
-
 Haloferax mediterranei (2R)-2-hydroxybutyrate + NAD+
-
 r
1.1.1.272 2-oxoisocaproate + NADH + H+
-
 Haloferax mediterranei 2-D-hydroxyisocaproate + NAD+
-
 r
1.1.1.272 2-oxoisoleucine + NAD(P)H
-
 Haloferax mediterranei 2-D-hydroxyisoleucine + NAD(P)+
-
 ir
1.1.1.272 2-oxoisoleucine + NAD(P)H
-
 Haloferax mediterranei R-4 / ATCC 33500 2-D-hydroxyisoleucine + NAD(P)+
-
 ir
1.1.1.272 2-oxoisoleucine + NADH + H+
-
 Haloferax mediterranei 2-D-hydroxyisoleucine + NAD+
-
 r
1.1.1.272 additional information no activity with 2-D-hydroxyacids and L-2-hydroxyisocaproic acid in oxidation reaction Haloferax mediterranei ?
-
 ?
1.1.1.272 additional information no activity with 2-D-hydroxyacids and L-2-hydroxyisocaproic acid in oxidation reaction Haloferax mediterranei R-4 / ATCC 33500 ?
-
 ?
1.1.1.272 pyruvate + NADPH + H+
-
 Haloferax mediterranei D-lactate + NADP+
-
 ir
1.1.1.272 pyruvate + NADPH + H+
-
 Haloferax mediterranei R-4 / ATCC 33500 D-lactate + NADP+
-
 ir

Subunits

EC Number Subunits Comment Organism
1.1.1.272 tetramer 4 * 33336, sequence calculation, 4 * 29500, recombinant enzyme, CTAB-PAGE, 4 * 47000, SDS-PAGE Haloferax mediterranei

Synonyms

EC Number Synonyms Comment Organism
1.1.1.272 2-D-hydroxyacid dehydrogenase
-
 Haloferax mediterranei
1.1.1.272 D-2-hydroxyacid dehydrogenase
-
 Haloferax mediterranei
1.1.1.272 D2-HDH
-
 Haloferax mediterranei

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.272 40
-
 assay at, recombinant refolded enzyme Haloferax mediterranei

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.272 5
-
 assay at, with substrate pyruvate Haloferax mediterranei
1.1.1.272 8.5
-
 assay at, all substrates, except for pyruvate Haloferax mediterranei

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.272 NADH
-
 Haloferax mediterranei
1.1.1.272 NADPH preferred cofactor Haloferax mediterranei