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Literature summary for 1.1.1.11 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

Activating Compound Comment Organism Structure
D-arabitol inducer Gluconobacter oxydans

Application

Application Comment Organism
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)

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

Inhibitors

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

KM Value [mM]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Ba2+ up to 200 mM, 2fold increase in activity Gluconobacter oxydans
Ca2+ up to 200 mM, 2fold increase in activity Gluconobacter oxydans
additional information not stimulatory: Mn2+, Ni2+, Mg2+ Gluconobacter oxydans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
28000
-
4 * 28500, calculated, 4 * 28000, SDS-PAGE Gluconobacter oxydans
28500
-
native aArDH, 2 * 28500, SDS-PAGE Gluconobacter oxydans
28500
-
recombinant aArDH, 2 * 28500, SDS-PAGE Gluconobacter oxydans
28500
-
4 * 28500, calculated, 4 * 28000, SDS-PAGE Gluconobacter oxydans
140000
-
gel filtration Gluconobacter oxydans

Organism

Organism UniProt Comment Textmining
Gluconobacter oxydans
-
-
-
Gluconobacter oxydans Q308C1
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Storage Stability

Storage Stability Organism
4°C, 100 mM Tris/HCl buffer, 2 mM 2-mercaptoethanol, 50 days Gluconobacter oxydans
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

Substrates and Products (Substrate)

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

Subunits

Subunits Comment Organism
heterodimer native aArDH, 2 * 28500, SDS-PAGE Gluconobacter oxydans
homodimer recombinant aArDH, 2 * 28500, SDS-PAGE Gluconobacter oxydans
tetramer 4 * 28500, calculated, 4 * 28000, SDS-PAGE Gluconobacter oxydans

Synonyms

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

Temperature Optimum [°C]

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

Temperature Range [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
optimum for ketone reduction Gluconobacter oxydans
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
8.5
-
optimum for polyol oxidation Gluconobacter oxydans

pH Stability

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

Cofactor

Cofactor Comment Organism Structure
NAD+ NAD+ is preferred over NADP+ Gluconobacter oxydans
NAD+ aArDH prefers NAD+ to NADP+ as coenzyme Gluconobacter oxydans
NADP+
-
Gluconobacter oxydans
NADP+ NAD+ is preferred over NADP+ Gluconobacter oxydans

pI Value

Organism Comment pI Value Maximum pI Value
Gluconobacter oxydans isoelectric focusing
-
5.88
Gluconobacter oxydans isoelectric focusing
-
5.9

Expression

Organism Comment Expression
Gluconobacter oxydans induced by D-arabinitol up

kcat/KM [mM/s]

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