Cloned (Comment) | Organism |
---|---|
recombinant overexpression of C-terminally His6-tagged enzyme in Escherichia coli | Ketogulonicigenium vulgare |
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, 20°C, X-ray diffraction structure determination and analysis at 1.64 A resolution, molecular replacement and modeling | Ketogulonicigenium vulgare |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-dehydro-L-gulonic acid + NADPH + H+ | Ketogulonicigenium vulgare | - |
L-idonate + NADP+ | - |
? | |
2-dehydro-L-gulonic acid + NADPH + H+ | Ketogulonicigenium vulgare Y25 | - |
L-idonate + NADP+ | - |
? | |
an (R)-2-hydroxycarboxylate + NADP+ | Ketogulonicigenium vulgare | - |
a 2-oxocarboxylate + NADPH + H+ | - |
? | |
an (R)-2-hydroxycarboxylate + NADP+ | Ketogulonicigenium vulgare Y25 | - |
a 2-oxocarboxylate + NADPH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ketogulonicigenium vulgare | - |
- |
- |
Ketogulonicigenium vulgare Y25 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli by nickel affinity and anion exchange chromatography, followed by gel filtration | Ketogulonicigenium vulgare |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-dehydro-L-gulonic acid + NADPH + H+ | - |
Ketogulonicigenium vulgare | L-idonate + NADP+ | - |
? | |
2-dehydro-L-gulonic acid + NADPH + H+ | - |
Ketogulonicigenium vulgare Y25 | L-idonate + NADP+ | - |
? | |
an (R)-2-hydroxycarboxylate + NADP+ | - |
Ketogulonicigenium vulgare | a 2-oxocarboxylate + NADPH + H+ | - |
? | |
an (R)-2-hydroxycarboxylate + NADP+ | - |
Ketogulonicigenium vulgare Y25 | a 2-oxocarboxylate + NADPH + H+ | - |
? |
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 flexibilty during catalysis | Ketogulonicigenium vulgare |
Synonyms | Comment | Organism |
---|---|---|
2-HDH | - |
Ketogulonicigenium vulgare |
HDH | - |
Ketogulonicigenium vulgare |
L-2-hydroxyacid dehydrogenase | - |
Ketogulonicigenium vulgare |
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 | |
NADP+ | - |
Ketogulonicigenium vulgare | |
NADPH | - |
Ketogulonicigenium vulgare |
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, cf. EC 1.1.1.215. The primary biochemical function of HDH from Ketogulonicigenium vulgare is C=O bond oxidation-reduction | Ketogulonicigenium vulgare |