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Literature summary extracted from

  • Jagtap, S.S.; Singh, R.; Kang, Y.C.; Zhao, H.; Lee, J.K.
    Cloning and characterization of a galactitol 2-dehydrogenase from Rhizobium legumenosarum and its application in D-tagatose production (2014), Enzyme Microb. Technol., 58-59, 44-51.
    View publication on PubMed

Application

EC Number Application Comment Organism
1.1.1.16 synthesis the enzyme can be used to produce optically pure building blocks and for the bioconversion of bioactive compounds Rhizobium leguminosarum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.16 gene rlgdh, recombinant overexpression of His-tagged enzyme in an active soluble form in Escherichia coli strain BL21 (DE3) Rhizobium leguminosarum

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.16 Fe2+
-
 Rhizobium leguminosarum
1.1.1.16 Zn2+
-
 Rhizobium leguminosarum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.16 additional information
-
 additional information Michaelis-Menten kinetics Rhizobium leguminosarum
1.1.1.16 8.8
-
 galactitol pH 9.5, 35°C Rhizobium leguminosarum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.16 Ba2+ activates Rhizobium leguminosarum
1.1.1.16 Ca2+ activates Rhizobium leguminosarum
1.1.1.16 Co2+ activates Rhizobium leguminosarum
1.1.1.16 Mg2+ required, the enzyme contains one magnesium atom per dimer, consistent with the presence of a structural metal ion. The homotetrameric enzyme contains two Mg2+ binding sites Rhizobium leguminosarum
1.1.1.16 Mn2+ activates Rhizobium leguminosarum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.16 28000
-
-
 Rhizobium leguminosarum
1.1.1.16 114000
-
 recombinant enzyme, gel filtration Rhizobium leguminosarum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.16 galactitol + NAD+ Rhizobium leguminosarum
-
 D-tagatose + NADH + H+
-
 ?
1.1.1.16 galactitol + NAD+ Rhizobium leguminosarum 3841
-
 D-tagatose + NADH + H+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.16 Rhizobium leguminosarum Q1MLL4 bv. viciae
-
1.1.1.16 Rhizobium leguminosarum 3841 Q1MLL4 bv. viciae
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.16 recombinant His-tagged enzyme in an active soluble form from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography Rhizobium leguminosarum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.16 1,2-butanediol + NAD+
-
 Rhizobium leguminosarum ? + NADH + H+
-
 ?
1.1.1.16 1,2-pentanediol + NAD+ high activity Rhizobium leguminosarum ? + NADH + H+
-
 ?
1.1.1.16 1,2-propanediol + NAD+
-
 Rhizobium leguminosarum ? + NADH + H+
-
 ?
1.1.1.16 1,6-hexanediol + NAD+ best substrate Rhizobium leguminosarum ? + NADH + H+
-
 ?
1.1.1.16 2-butanol + NAD+
-
 Rhizobium leguminosarum ? + NADH + H+
-
 ?
1.1.1.16 adonitol + NAD+
-
 Rhizobium leguminosarum ? + NADH + H+
-
 ?
1.1.1.16 D-arabinitol + NAD+
-
 Rhizobium leguminosarum ? + NADH + H+
-
 ?
1.1.1.16 D-mannitol + NAD+
-
 Rhizobium leguminosarum ? + NADH + H+
-
 ?
1.1.1.16 galactitol + NAD+
-
 Rhizobium leguminosarum D-tagatose + NADH + H+
-
 ?
1.1.1.16 galactitol + NAD+ high activity Rhizobium leguminosarum D-tagatose + NADH + H+
-
 ?
1.1.1.16 galactitol + NAD+
-
 Rhizobium leguminosarum 3841 D-tagatose + NADH + H+
-
 ?
1.1.1.16 galactitol + NAD+ high activity Rhizobium leguminosarum 3841 D-tagatose + NADH + H+
-
 ?
1.1.1.16 L-arabinitol + NAD+
-
 Rhizobium leguminosarum L-ribulose + NADH + H+
-
 ?
1.1.1.16 L-arabinitol + NAD+
-
 Rhizobium leguminosarum 3841 L-ribulose + NADH + H+
-
 ?
1.1.1.16 additional information the purified recombinant RlGDH catalyzes the oxidation of a wide range of substrates, including polyvalent aliphatic alcohols and polyols, to the correspondingketones and ketoses. Among various polyols, galactitol is the preferred substrate of the enzyme. No activity with ethanol, pentanol, and propanol Rhizobium leguminosarum ?
-
 ?
1.1.1.16 additional information the purified recombinant RlGDH catalyzes the oxidation of a wide range of substrates, including polyvalent aliphatic alcohols and polyols, to the correspondingketones and ketoses. Among various polyols, galactitol is the preferred substrate of the enzyme. No activity with ethanol, pentanol, and propanol Rhizobium leguminosarum 3841 ?
-
 ?
1.1.1.16 myosinitol + NAD+
-
 Rhizobium leguminosarum ? + NADH + H+
-
 ?
1.1.1.16 sorbitol + NAD+
-
 Rhizobium leguminosarum D-fructose + NADH + H+
-
 ?
1.1.1.16 xylitol + NAD+
-
 Rhizobium leguminosarum L-xylulose + NADH + H+
-
 ?
1.1.1.16 xylitol + NAD+
-
 Rhizobium leguminosarum 3841 L-xylulose + NADH + H+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.16 homotetramer 4 * 28000, recombinant enzyme, SDS-PAGE, 4 * 27289, sequence calculation Rhizobium leguminosarum
1.1.1.16 More sequence alignment and secondary structure prediction, homology modeling and sequence analysis, overview Rhizobium leguminosarum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.16 35
-
 recombinant enzyme Rhizobium leguminosarum

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.16 15 55 activity profile, overview Rhizobium leguminosarum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.16 30 40 when tested at 30°C and 35°C, the enzyme is able to retain about 50% of its activity after 114 and 54 min, respectively, while at 40°C, 50% of its activity is retained after 28 min Rhizobium leguminosarum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.16 13.91
-
 galactitol pH 9.5, 35°C Rhizobium leguminosarum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.16 9.5
-
 recombinant enzyme Rhizobium leguminosarum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.16 6.5 10 activity profile, overview. The enzyme shows over 60% of its maximum activity at pH values of 8.0-10.0 Rhizobium leguminosarum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.16 additional information the cofactor binding site in short-chain dehydrogenases/reductases enzymes is characterized by the conserved NAD-binding glycine-rich Rossmann fold domain (TGGGRAIG) Rhizobium leguminosarum
1.1.1.16 NAD+
-
 Rhizobium leguminosarum

General Information

EC Number General Information Comment Organism
1.1.1.16 evolution galactitol 2-dehydrogenase belongs to the protein subfamily of short-chain dehydrogenases/reductases Rhizobium leguminosarum
1.1.1.16 additional information homology modeling and sequence analysisA , overview. The active site contains residues Asn119,Ser147, Tyr162, and Lys166 which are conserved in short-chain dehydrogenases/reductases Rhizobium leguminosarum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.16 1.581
-
 galactitol pH 9.5, 35°C Rhizobium leguminosarum