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Literature summary for 1.1.1.215 extracted from

  • Han, X.; Xiong, X.; Hu, X.; Li, M.; Zhang, W.; Liu, X.
    Crystallization and structural analysis of 2-hydroxyacid dehydrogenase from Ketogulonicigenium vulgare (2014), Biotechnol. Lett., 36, 295-300 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
recombinant overexpression of C-terminally His6-tagged enzyme in Escherichia coli Ketogulonicigenium vulgare

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant enzyme, sitting drop vapour diffusion method, mixing of 0.001 ml of 30 mg/ml protein in 25 mM Tris/HCl, pH 8.0, 0.3 M NaCl, and 1 mM dithiothreitol, with 0.001 ml of reservoir solution containing 23% w/v PEG 3350, 0.2 M MgCl2, and 0.1 M HEPES, pH 7.0, and equilibration against 0.1 ml of reservoir solution, 20Ā°C, X-ray diffraction structure determination and analysis at 1.64 A resolution, molecular replacement and modeling Ketogulonicigenium vulgare

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05
-
NADPH recombinant His6-tagged enzyme, pH and temperature not specified in the publication Ketogulonicigenium vulgare
0.11
-
NADH recombinant His6-tagged enzyme, pH and temperature not specified in the publication Ketogulonicigenium vulgare
2.6
-
2-keto-L-gluconic acid recombinant His6-tagged enzyme, pH and temperature not specified in the publication Ketogulonicigenium vulgare

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-keto-L-gluconic acid + NADPH + H+ Ketogulonicigenium vulgare
-
L-idonate + NADP+
-
?
2-keto-L-gluconic acid + NADPH + H+ Ketogulonicigenium vulgare Y25
-
L-idonate + NADP+
-
?

Organism

Organism UniProt Comment Textmining
Ketogulonicigenium vulgare
-
-
-
Ketogulonicigenium vulgare Y25
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli by nickel affinity and anion exchange chromatography, followed by gel filtration Ketogulonicigenium vulgare

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-keto-L-gluconic acid + NADH + H+
-
Ketogulonicigenium vulgare L-idonate + NAD+
-
?
2-keto-L-gluconic acid + NADH + H+
-
Ketogulonicigenium vulgare Y25 L-idonate + NAD+
-
?
2-keto-L-gluconic acid + NADPH + H+
-
Ketogulonicigenium vulgare L-idonate + NADP+
-
?
2-keto-L-gluconic acid + NADPH + H+
-
Ketogulonicigenium vulgare Y25 L-idonate + NADP+
-
?

Subunits

Subunits Comment Organism
More the enzyme shows a structure consisting of two-compact domains separated by a deep active cleft. This typical topology is conserved in other 2-HDH. The smaller domain is the substrate binding domain or catalytic domain, which is formed from N-terminal residues 2-102 and C-terminal residues 288-317. It is folded into a five-stranded parallel beta-sheet flanked by five alpha-helices, forming a modified Rossmann topology. The larger domain is responsible for binding the cofactor and contains a conserved [GXGXXG (X17) D] motif that is characteristic of the NAD(P)H/NAD(P)+-binding region. It consists of residues 103-287, forming a seven-stranded parallel beta-sheet flanked by seven alpha-helices. A two-stranded hinge connects the two domains showing flexibility during catalysis Ketogulonicigenium vulgare

Synonyms

Synonyms Comment Organism
2-HDH
-
Ketogulonicigenium vulgare
HDH
-
Ketogulonicigenium vulgare
L-2-hydroxyacid dehydrogenase
-
Ketogulonicigenium vulgare

Cofactor

Cofactor Comment Organism Structure
additional information the larger enzyme domain is responsible for binding the cofactor and contains a conserved [GXGXXG (X17) D] motif that is characteristic of the NAD(P)H/NAD(P)+-binding region. It consists of residues 103-287, forming a seven-stranded parallel beta-sheet flanked by seven alpha-helices Ketogulonicigenium vulgare
NADH
-
Ketogulonicigenium vulgare
NADPH
-
Ketogulonicigenium vulgare

General Information

General Information Comment Organism
additional information the amino acid residues Arg234, Glu263 and His 279 form the active site of enzyme HDH. Residues Arg234, Ala210, Thr211, and Arg212, which are located on top of the catalytic triad, act as a size filter to jointly determine the substrate specificity Ketogulonicigenium vulgare
physiological function the enzyme catalyzes the bioconversion of 2-dehydro-L-gulonic acid to L-idonate, which plays a negative role in the manufacture of vitamin C. The primary biochemical function of HDH from Ketogulonicigenium vulgare is C=O bond oxidation-reduction, cf. EC 1.1.1.272 Ketogulonicigenium vulgare