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Literature summary extracted from

  • Fredslund, F.; Otten, H.; Gemperlein, S.; Poulsen, J.C.; Carius, Y.; Kohring, G.W.; Lo Leggio, L.
    Structural characterization of the thermostable Bradyrhizobium japonicum D-sorbitol dehydrogenase (2016), Acta Crystallogr. Sect. F, 72, 846-852.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.15 gene rdh, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21-Gold (DE3) Bradyrhizobium japonicum
1.1.99.21 expressed in Escherichia coli BL21-Gold (DE3) cells Bradyrhizobium japonicum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.15 purified recombinant His-tagged enzyme in complex with both NAD+ and glucitol, X-ray diffraction structure determination and analysis at 2.9 A resolution, molecular replacement and modelling Bradyrhizobium japonicum
1.1.99.21 sitting drop vapor diffusion method, using 1.4 M NaH2PO4/K2HPO4 pH 5.6 Bradyrhizobium japonicum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.15 Zn2+ zinc metalloenzyme Bradyrhizobium japonicum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.15 26900
-
 SDS-PAGE, subunit Bradyrhizobium japonicum
1.1.1.15 77700
-
 native PAGE, recombinant enzyme Bradyrhizobium japonicum
1.1.1.15 83200
-
 gel filtration, recombinant enzyme Bradyrhizobium japonicum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.15 D-iditol + NAD+ Bradyrhizobium japonicum i.e. D-sorbitol D-sorbose + NADH + H+
-
 r
1.1.1.15 D-iditol + NAD+ Bradyrhizobium japonicum USDA110 i.e. D-sorbitol D-sorbose + NADH + H+
-
 r
1.1.99.21 D-glucitol + NAD+ Bradyrhizobium japonicum
-
 D-fructose + NADH + H+
-
 ?
1.1.99.21 D-glucitol + NAD+ Bradyrhizobium japonicum USDA110
-
 D-fructose + NADH + H+
-
 ?
1.1.99.21 L-glucitol + NAD+ Bradyrhizobium japonicum 90% conversion D-sorbose + NADH + H+
-
 ?
1.1.99.21 L-glucitol + NAD+ Bradyrhizobium japonicum USDA110 90% conversion D-sorbose + NADH + H+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.15 Bradyrhizobium japonicum Q89FN7 i.e. Bradyrhizobium diazoefficiens
-
1.1.1.15 Bradyrhizobium japonicum USDA110 Q89FN7 i.e. Bradyrhizobium diazoefficiens
-
1.1.99.21 Bradyrhizobium japonicum
-
-
-
1.1.99.21 Bradyrhizobium japonicum USDA110
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.15 recombinant His-tagged enzyme from Escherichia coli strain BL21-Gold (DE3) by heat treatment at 50°C and metal affinity chromatography to homogeneity Bradyrhizobium japonicum
1.1.99.21 metal affinity column chromatography Bradyrhizobium japonicum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.15 D-glucitol + NAD+
-
 Bradyrhizobium japonicum D-fructose + NADH + H+
-
 r
1.1.1.15 D-glucitol + NAD+
-
 Bradyrhizobium japonicum USDA110 D-fructose + NADH + H+
-
 r
1.1.1.15 D-iditol + NAD+ i.e. D-sorbitol Bradyrhizobium japonicum D-sorbose + NADH + H+
-
 r
1.1.1.15 D-iditol + NAD+ i.e D-sorbitol Bradyrhizobium japonicum D-sorbose + NADH + H+
-
 r
1.1.1.15 D-iditol + NAD+ i.e. D-sorbitol Bradyrhizobium japonicum USDA110 D-sorbose + NADH + H+
-
 r
1.1.1.15 D-iditol + NAD+ i.e D-sorbitol Bradyrhizobium japonicum USDA110 D-sorbose + NADH + H+
-
 r
1.1.1.15 L-glucitol + NAD+ i.e. D-sorbitol, 90% conversion of L-glucitol to D-sorbose Bradyrhizobium japonicum D-sorbose + NADH + H+
-
 r
1.1.1.15 L-glucitol + NAD+ i.e. D-sorbitol, 90% conversion of L-glucitol to D-sorbose Bradyrhizobium japonicum USDA110 D-sorbose + NADH + H+
-
 r
1.1.1.15 additional information the substrate in BjSDH is stabilized via the side chain of Glu150, Ser140 and the carbonyl groups of Pro183 and Gly184 Bradyrhizobium japonicum ?
-
 ?
1.1.1.15 additional information the substrate in BjSDH is stabilized via the side chain of Glu150, Ser140 and the carbonyl groups of Pro183 and Gly184 Bradyrhizobium japonicum USDA110 ?
-
 ?
1.1.99.21 D-glucitol + NAD+
-
 Bradyrhizobium japonicum D-fructose + NADH + H+
-
 ?
1.1.99.21 D-glucitol + NAD+
-
 Bradyrhizobium japonicum USDA110 D-fructose + NADH + H+
-
 ?
1.1.99.21 L-glucitol + NAD+ 90% conversion Bradyrhizobium japonicum D-sorbose + NADH + H+
-
 ?
1.1.99.21 L-glucitol + NAD+ 90% conversion Bradyrhizobium japonicum USDA110 D-sorbose + NADH + H+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.15 More a continuous beta-sheet is formed between two monomers in the tetramer. The BjSDH structure folds in the Rossmann fold. The monomer of BjSDH is composed of a central parallel beta-sheet surrounded by three alpha-helices on each side. At the top of the beta-sheet a small helix–turn–helix motif is located between the two final beta-strands betaF and betaG. Tetrameric quaternary organization, structure comparisons, overview Bradyrhizobium japonicum
1.1.1.15 tetramer 4 * 26900, SDS-PAGE and crystal structure analysis Bradyrhizobium japonicum

Synonyms

EC Number Synonyms Comment Organism
1.1.1.15 BjSDH
-
 Bradyrhizobium japonicum
1.1.1.15 D-sorbitol dehydrogenase
-
 Bradyrhizobium japonicum
1.1.99.21 SDH
-
 Bradyrhizobium japonicum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.99.21 47
-
 melting temperature Bradyrhizobium japonicum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.15 NAD+
-
 Bradyrhizobium japonicum
1.1.1.15 NADH dependent on Bradyrhizobium japonicum
1.1.99.21 NAD+
-
 Bradyrhizobium japonicum

General Information

EC Number General Information Comment Organism
1.1.1.15 evolution the enzyme BjSDH is part of the superfamily of Zn-independent short-chain dehydrogenases. The BjSDH structure folds in the Rossmann fold, which is typical for NADH-dependent enzymes and the SDR family Bradyrhizobium japonicum