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Literature summary extracted from

  • Connaris, H.; Chaudhuri, J.B.; Danson, M.J.; Hough, D.W.
    Expression, reactivation, and purification of enzymes from Haloferax volcanii in Escherichia coli. (1999), Biotechnol. Bioeng., 64, 38-45.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.8.1.4 expressed in Escherichia coli using the cytoplasmic expression vectors, pET3a and pET3d. Expressed as inclusion bodies and refolded by solubilisation in 8 M urea followed by dilution into a buffer containing 2 M KCl, 0.010 mM FAD, 1 mM NAD+, and 0.3 mM GSSG/3 mM GSH Haloferax volcanii
2.3.3.16 expressed in Escherichia coli using the cytoplasmic expression vectors, pET3a and pET3d. The inactive enzyme is reactivated following overnight incubation in 2 M KCl Haloferax volcanii

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.3.16 KCl at 0.2 M KCl, approximately 60% of its maximal activity Haloferax volcanii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.8.1.4 0.046
-
 dihydrolipoamide 37°C, pH 8.0 Haloferax volcanii
1.8.1.4 0.156
-
 NAD+ 37°C, pH 8.0 Haloferax volcanii
2.3.3.16 0.026
-
 oxaloacetate 30°C, pH 8.0, native enzyme Haloferax volcanii
2.3.3.16 0.027
-
 oxaloacetate 30°C, pH 8.0, recombinant enzyme Haloferax volcanii
2.3.3.16 123
-
 acetyl-CoA 30°C, pH 8.0, recombinant enzyme Haloferax volcanii
2.3.3.16 125
-
 acetyl-CoA 30°C, pH 8.0, native enzyme Haloferax volcanii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.3.16 42000
-
 x * 42000, SDS-PAGE Haloferax volcanii

Organism

EC Number Organism UniProt Comment Textmining
1.8.1.4 Haloferax volcanii Q04829
-
-
1.8.1.4 Haloferax volcanii DSM 3757 Q04829
-
-
2.3.3.16 Haloferax volcanii D4GS06
-
-
2.3.3.16 Haloferax volcanii DSM 3757 D4GS06
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.8.1.4 purified using copper-based metal ion affinity chromatography in the presence of 2 M KCl Haloferax volcanii
2.3.3.16 recovered using dye-affinity chromatography in the presence of salt Haloferax volcanii

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.8.1.4 20
-
 37°C, pH 8.0 Haloferax volcanii
2.3.3.16 40
-
 37°C, pH 8.0 Haloferax volcanii

Storage Stability

EC Number Storage Stability Organism
2.3.3.16 room temperature, at 0.2 M KCl, the enzyme is stable at for at least 48 h Haloferax volcanii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.8.1.4 dihydrolipoamide + NAD+
-
 Haloferax volcanii lipoamide + NADH
-
 ?
1.8.1.4 dihydrolipoamide + NAD+
-
 Haloferax volcanii DSM 3757 lipoamide + NADH
-
 ?
2.3.3.16 acetyl-CoA + H2O + oxaloacetate
-
 Haloferax volcanii citrate + CoA
-
 ?
2.3.3.16 acetyl-CoA + H2O + oxaloacetate
-
 Haloferax volcanii DSM 3757 citrate + CoA
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.3.16 ? x * 42000, SDS-PAGE Haloferax volcanii

Synonyms

EC Number Synonyms Comment Organism
1.8.1.4 DHLipDH
-
 Haloferax volcanii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.8.1.4 37
-
 assay at Haloferax volcanii
2.3.3.16 30
-
 assay at Haloferax volcanii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.8.1.4 8
-
 assay at Haloferax volcanii
2.3.3.16 8
-
 assay at Haloferax volcanii