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Literature summary for 1.1.1.16 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

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

Cloned(Commentary)

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

Inhibitors

Inhibitors Comment Organism Structure
Fe2+
-
Rhizobium leguminosarum
Zn2+
-
Rhizobium leguminosarum

KM Value [mM]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Ba2+ activates Rhizobium leguminosarum
Ca2+ activates Rhizobium leguminosarum
Co2+ activates Rhizobium leguminosarum
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
Mn2+ activates Rhizobium leguminosarum

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

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

Purification (Commentary)

Purification (Comment) Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1,2-butanediol + NAD+
-
Rhizobium leguminosarum ? + NADH + H+
-
?
1,2-pentanediol + NAD+ high activity Rhizobium leguminosarum ? + NADH + H+
-
?
1,2-propanediol + NAD+
-
Rhizobium leguminosarum ? + NADH + H+
-
?
1,6-hexanediol + NAD+ best substrate Rhizobium leguminosarum ? + NADH + H+
-
?
2-butanol + NAD+
-
Rhizobium leguminosarum ? + NADH + H+
-
?
adonitol + NAD+
-
Rhizobium leguminosarum ? + NADH + H+
-
?
D-arabinitol + NAD+
-
Rhizobium leguminosarum ? + NADH + H+
-
?
D-mannitol + NAD+
-
Rhizobium leguminosarum ? + NADH + H+
-
?
galactitol + NAD+
-
Rhizobium leguminosarum D-tagatose + NADH + H+
-
?
galactitol + NAD+ high activity Rhizobium leguminosarum D-tagatose + NADH + H+
-
?
galactitol + NAD+
-
Rhizobium leguminosarum 3841 D-tagatose + NADH + H+
-
?
galactitol + NAD+ high activity Rhizobium leguminosarum 3841 D-tagatose + NADH + H+
-
?
L-arabinitol + NAD+
-
Rhizobium leguminosarum L-ribulose + NADH + H+
-
?
L-arabinitol + NAD+
-
Rhizobium leguminosarum 3841 L-ribulose + NADH + H+
-
?
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 ?
-
?
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 ?
-
?
myosinitol + NAD+
-
Rhizobium leguminosarum ? + NADH + H+
-
?
sorbitol + NAD+
-
Rhizobium leguminosarum D-fructose + NADH + H+
-
?
xylitol + NAD+
-
Rhizobium leguminosarum L-xylulose + NADH + H+
-
?
xylitol + NAD+
-
Rhizobium leguminosarum 3841 L-xylulose + NADH + H+
-
?

Subunits

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

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
35
-
recombinant enzyme Rhizobium leguminosarum

Temperature Range [┬░C]

Temperature Minimum [┬░C] Temperature Maximum [┬░C] Comment Organism
15 55 activity profile, overview Rhizobium leguminosarum

Temperature Stability [┬░C]

Temperature Stability Minimum [┬░C] Temperature Stability Maximum [┬░C] Comment Organism
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]

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

pH Optimum

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

pH Range

pH Minimum pH Maximum Comment Organism
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

Cofactor Comment Organism Structure
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
NAD+
-
Rhizobium leguminosarum

General Information

General Information Comment Organism
evolution galactitol 2-dehydrogenase belongs to the protein subfamily of short-chain dehydrogenases/reductases Rhizobium leguminosarum
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]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.581
-
galactitol pH 9.5, 35┬░C Rhizobium leguminosarum