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Literature summary extracted from

  • Cheng, H.; Li, Z.; Jiang, N.; Deng, Z.
    Cloning, purification and characterization of an NAD-dependent D-arabitol dehydrogenase from acetic acid bacterium, Acetobacter suboxydans (2009), Protein J., 28, 263-272.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.11 D-arabitol inducer Gluconobacter oxydans

Application

EC Number Application Comment Organism
1.1.1.11 industry promising method for the production of xylitol from the cheap material glucose if the aArDH gene can be introduced into yeast strains that can convert glucose to D-arabitol Gluconobacter oxydans

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.11 expression in Escherichia coli Gluconobacter oxydans
1.1.1.11 expression vector containing the ORF of aArDH under the control of T7 promoter, pET21aArDH, transformed into Escherichia coli strain BL21(DE3) Gluconobacter oxydans
1.1.1.287 expression in Escherichia coli Gluconobacter oxydans

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.11 Cu2+ 10 mM, complete inhibition. Activity can partly be restored by addition of EDTA Gluconobacter oxydans
1.1.1.11 CuSO4 inactivates aArDH activity, which is restored by 80% by addition of 100 mM EDTA at pH 8.0 Gluconobacter oxydans
1.1.1.11 Zn2+ up to 200 mM, slight inhibition Gluconobacter oxydans
1.1.1.287 Cu2+ 10 mM, complete inhibition. Up to 80% renaturation by 100 mM EDTA Gluconobacter oxydans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.11 4.5
-
 D-arabitol
-
 Gluconobacter oxydans
1.1.1.11 4.5
-
 D-arabinitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 18.5
-
 xylitol
-
 Gluconobacter oxydans
1.1.1.11 78.5
-
 D-mannitol
-
 Gluconobacter oxydans
1.1.1.11 78.5
-
 D-mannitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 127.3
-
 D-sorbitol
-
 Gluconobacter oxydans
1.1.1.11 127.3
-
 D-sorbitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 133.6
-
 glycerol
-
 Gluconobacter oxydans
1.1.1.11 133.6
-
 glycerol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 177.2
-
 xylitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 242.8
-
 ribitol
-
 Gluconobacter oxydans
1.1.1.11 242.8
-
 ribitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 4.5
-
 D-arabitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 18.5
-
 xylitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 78.5
-
 D-mannitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 127.3
-
 D-sorbitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 133.6
-
 glycerol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 242.8
-
 D-ribitol pH 8.5, 25°C Gluconobacter oxydans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.11 Ba2+ up to 200 mM, 2fold increase in activity Gluconobacter oxydans
1.1.1.11 Ca2+ up to 200 mM, 2fold increase in activity Gluconobacter oxydans
1.1.1.11 additional information not stimulatory: Mn2+, Ni2+, Mg2+ Gluconobacter oxydans
1.1.1.287 Ba2+ up to 200 mM, 2fold increase in activity Gluconobacter oxydans
1.1.1.287 Ca2+ up to 200 mM, 2fold increase in activity Gluconobacter oxydans
1.1.1.287 additional information no effect by addition of Mn2+, Ni2+, Mg2+ Gluconobacter oxydans

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.11 28000
-
 4 * 28500, calculated, 4 * 28000, SDS-PAGE Gluconobacter oxydans
1.1.1.11 28500
-
 native aArDH, 2 * 28500, SDS-PAGE Gluconobacter oxydans
1.1.1.11 28500
-
 recombinant aArDH, 2 * 28500, SDS-PAGE Gluconobacter oxydans
1.1.1.11 28500
-
 4 * 28500, calculated, 4 * 28000, SDS-PAGE Gluconobacter oxydans
1.1.1.11 140000
-
 gel filtration Gluconobacter oxydans
1.1.1.287 28500
-
 2 * 28500, SDS-PAGE and calculated, recombinant enzyme Gluconobacter oxydans
1.1.1.287 28500
-
 4 * 28500, SDS-PAGE and calculated, native enzyme Gluconobacter oxydans
1.1.1.287 70000
-
 gel filtration, recombinant enzyme Gluconobacter oxydans
1.1.1.287 140000
-
 gel filtration, native enzyme Gluconobacter oxydans

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.11 Gluconobacter oxydans
-
-
-
1.1.1.11 Gluconobacter oxydans Q308C1
-
-
1.1.1.287 Gluconobacter oxydans Q308C1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.11
-
 Gluconobacter oxydans
1.1.1.11 recombinant and native enzymes purified to homogeneity by ammonium sulfate precipitation and gel filtration, 119fold Gluconobacter oxydans
1.1.1.287 native enzyme Gluconobacter oxydans

Renatured (Commentary)

EC Number Renatured (Comment) Organism
1.1.1.287 inhibition by Cu2+ can partly be restored by EDTA Gluconobacter oxydans

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.11 0.14
-
 crude extract Gluconobacter oxydans
1.1.1.11 0.55
-
 recombinant aArDH with meso-erythritol as substrate Gluconobacter oxydans
1.1.1.11 0.87
-
 recombinant aArDH with ethanol as substrate Gluconobacter oxydans
1.1.1.11 1.5
-
 recombinant aArDH with ribitol as substrate Gluconobacter oxydans
1.1.1.11 2.1
-
 recombinant aArDH with glycerol as substrate Gluconobacter oxydans
1.1.1.11 2.5
-
 recombinant aArDH with D-fructose as substrate Gluconobacter oxydans
1.1.1.11 2.8
-
 recombinant aArDH with D-sorbitol as substrate Gluconobacter oxydans
1.1.1.11 4.2
-
 recombinant aArDH with D-mannitol as substrate Gluconobacter oxydans
1.1.1.11 12.6
-
 recombinant aArDH with xylitol as substrate Gluconobacter oxydans
1.1.1.11 16.4
-
 recombinant aArDH with D-xylulose as substrate Gluconobacter oxydans
1.1.1.11 16.66
-
 119fold purified enzyme Gluconobacter oxydans
1.1.1.11 68.5
-
 pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 68.5
-
 recombinant aArDH with 100 mM D-arabitol as substrate Gluconobacter oxydans
1.1.1.287 12.6
-
 substrate D-xylitol, pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 16.4
-
 substrate D-xylulose, pH 5.5, 25°C Gluconobacter oxydans
1.1.1.287 68.5
-
 substrate D-arabitol, pH 8.5, 25°C Gluconobacter oxydans

Storage Stability

EC Number Storage Stability Organism
1.1.1.11 4°C, 100 mM Tris/HCl buffer, 2 mM 2-mercaptoethanol, 50 days Gluconobacter oxydans
1.1.1.11 4°C, 100 mM Tris/HCl with 2 mM 2-mercaptoethanol for 50 days, 50% loss of activity. Addition of 2-mercaptoethanol is required Gluconobacter oxydans
1.1.1.287 4°C, 100 mM Tris/HCl with 2 mM 2-mercaptoethanol, 50 days, 50% oxidative activity Gluconobacter oxydans
1.1.1.287 4°C, 100 mM Tris/HCl with 2 mM 2-mercaptoethanol, 80 days, 5% oxidative activity Gluconobacter oxydans
1.1.1.287 4°C, 100 mM Tris/HCl without 2 mM 2-mercaptoethanol, 20 days, complete loss of activity Gluconobacter oxydans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.11 D-arabinitol + NAD+
-
 Gluconobacter oxydans D-xylulose + NADH + H+
-
 r
1.1.1.11 D-arabitol + NAD+ is the optimal substrate for aArDH Gluconobacter oxydans D-xylulose + NADH + H+
-
 r
1.1.1.11 D-fructose + NADH + H+
-
 Gluconobacter oxydans D-mannitol + NAD+
-
 ?
1.1.1.11 D-mannitol + NAD+
-
 Gluconobacter oxydans D-fructose + NADH + H+
-
 ?
1.1.1.11 D-sorbitol + NAD+
-
 Gluconobacter oxydans L-sorbose + NADH + H+
-
 ?
1.1.1.11 D-xylulose + NADH + H+
-
 Gluconobacter oxydans D-arabitol + NAD+
-
 r
1.1.1.11 ethanol + NAD+
-
 Gluconobacter oxydans acetaldehyde + NADH + H+
-
 ?
1.1.1.11 glycerol + NAD+
-
 Gluconobacter oxydans dihydroxyacetone + NADH + H+
-
 ?
1.1.1.11 meso-erythritol + NAD+
-
 Gluconobacter oxydans ? + NADH + H+
-
 ?
1.1.1.11 additional information both purified native and recombinant aArDH do not accept L-arabitol as substrate Gluconobacter oxydans ?
-
 ?
1.1.1.11 additional information no substrate: L-arabinitol Gluconobacter oxydans ?
-
 ?
1.1.1.11 ribitol + NAD+
-
 Gluconobacter oxydans D-ribulose + NADH + H+
-
 ?
1.1.1.11 xylitol + NAD+
-
 Gluconobacter oxydans D-xylulose + NADH + H+
-
 ?
1.1.1.287 D-arabitol + NAD+
-
 Gluconobacter oxydans D-xylulose + NADH + H+
-
 ?
1.1.1.287 D-arabitol + NADP+
-
 Gluconobacter oxydans D-xylulose + NADPH + H+
-
 r
1.1.1.287 D-mannitol + NAD+
-
 Gluconobacter oxydans D-fructose + NADH + H+
-
 ?
1.1.1.287 D-ribitol + NAD+
-
 Gluconobacter oxydans D-ribulose + NADH + H+
-
 ?
1.1.1.287 D-sorbitol + NAD+
-
 Gluconobacter oxydans sorbose + NADH + H+
-
 ?
1.1.1.287 D-xylulose + NAD+
-
 Gluconobacter oxydans D-arabitol + NADH + H+
-
 r
1.1.1.287 glycerol + NAD+
-
 Gluconobacter oxydans dihydroxyacetone + NADH + H+
-
 ?
1.1.1.287 xylitol + NAD+
-
 Gluconobacter oxydans D-xylulose + NADH + H+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.11 heterodimer native aArDH, 2 * 28500, SDS-PAGE Gluconobacter oxydans
1.1.1.11 homodimer recombinant aArDH, 2 * 28500, SDS-PAGE Gluconobacter oxydans
1.1.1.11 tetramer 4 * 28500, calculated, 4 * 28000, SDS-PAGE Gluconobacter oxydans
1.1.1.287 dimer 2 * 28500, SDS-PAGE and calculated, recombinant enzyme Gluconobacter oxydans
1.1.1.287 tetramer 4 * 28500, SDS-PAGE and calculated, native enzyme Gluconobacter oxydans

Synonyms

EC Number Synonyms Comment Organism
1.1.1.11 aArDH
-
 Gluconobacter oxydans
1.1.1.11 D-xylulose-forming D-arabitol dehydrogenase
-
 Gluconobacter oxydans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.11 30
-
 optimal temperature for oxidation is at 30°C, with 2% of the maximum activity at 50°C Gluconobacter oxydans
1.1.1.287 30
-
-
 Gluconobacter oxydans

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.11 30 50 optimal temperature for oxidation is at 30°C, with 2% of the maximum activity at 50°C Gluconobacter oxydans
1.1.1.287 50
-
 2% of maximum activity Gluconobacter oxydans

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.11 13.4
-
 ribitol
-
 Gluconobacter oxydans
1.1.1.11 13.4
-
 ribitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 16.3
-
 glycerol
-
 Gluconobacter oxydans
1.1.1.11 16.3
-
 glycerol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 19.7
-
 D-sorbitol
-
 Gluconobacter oxydans
1.1.1.11 19.7
-
 D-sorbitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 35.3
-
 D-mannitol
-
 Gluconobacter oxydans
1.1.1.11 35.3
-
 D-mannitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 177.2
-
 xylitol
-
 Gluconobacter oxydans
1.1.1.11 177.2
-
 xylitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 876
-
 D-arabitol
-
 Gluconobacter oxydans
1.1.1.11 876
-
 D-arabinitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 13.4
-
 D-ribitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 16.3
-
 glycerol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 19.7
-
 D-sorbitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 35.3
-
 D-mannitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 177.2
-
 xylitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 876
-
 D-arabitol pH 8.5, 25°C Gluconobacter oxydans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.11 5.5
-
 optimum for ketone reduction Gluconobacter oxydans
1.1.1.11 8.5
-
 optimum pH for oxidation is 8.5, with 0.7% and 5.6% of the maximum activity at pH 5.0 and 14, respectively. Optimal pH for reduction is 5.5, with 2 and 10% of the maximum activity at pH 4.5 and 8.0 Gluconobacter oxydans
1.1.1.11 8.5
-
 optimum for polyol oxidation Gluconobacter oxydans

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.287 5.5
-
 ketone reduction Gluconobacter oxydans
1.1.1.287 8.5
-
 polyol oxidation Gluconobacter oxydans

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.1.1.11 8.5 12 very stable in alkaline buffer Gluconobacter oxydans

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.11 NAD+ NAD+ is preferred over NADP+ Gluconobacter oxydans
1.1.1.11 NAD+ aArDH prefers NAD+ to NADP+ as coenzyme Gluconobacter oxydans
1.1.1.11 NADP+
-
 Gluconobacter oxydans
1.1.1.11 NADP+ NAD+ is preferred over NADP+ Gluconobacter oxydans
1.1.1.287 NADH NADH is preferred over NADPH Gluconobacter oxydans
1.1.1.287 NADPH NADH is preferred over NADPH Gluconobacter oxydans

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.1.1.11 Gluconobacter oxydans isoelectric focusing
-
 5.88
1.1.1.11 Gluconobacter oxydans isoelectric focusing
-
 5.9
1.1.1.287 Gluconobacter oxydans isoelectric focusing
-
 5.9

Expression

EC Number Organism Comment Expression
1.1.1.11 Gluconobacter oxydans induced by D-arabinitol up
1.1.1.287 Gluconobacter oxydans induced by growth on D-arabinitol up

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.11 0.06
-
 ribitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 0.12
-
 glycerol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 0.15
-
 D-sorbitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 0.45
-
 D-mannitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 9.58
-
 xylitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.11 194.7
-
 D-arabinitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 0.06
-
 D-ribitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 0.12
-
 glycerol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 0.15
-
 D-sorbitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 0.45
-
 D-mannitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 9.58
-
 xylitol pH 8.5, 25°C Gluconobacter oxydans
1.1.1.287 194.7
-
 D-arabitol pH 8.5, 25°C Gluconobacter oxydans