Activating Compound | Comment | Organism | Structure |
---|---|---|---|
D-arabitol | inducer | Gluconobacter oxydans |
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 (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 | 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 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 | 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Gluconobacter oxydans | - |
- |
- |
Gluconobacter oxydans | Q308C1 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Gluconobacter oxydans |
recombinant and native enzymes purified to homogeneity by ammonium sulfate precipitation and gel filtration, 119fold | Gluconobacter oxydans |
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 | 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 | 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 | 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 | Comment | Organism |
---|---|---|
aArDH | - |
Gluconobacter oxydans |
D-xylulose-forming D-arabitol dehydrogenase | - |
Gluconobacter oxydans |
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 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 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 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 Maximum | Comment | Organism |
---|---|---|---|
8.5 | 12 | very stable in alkaline buffer | Gluconobacter oxydans |
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 |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Gluconobacter oxydans | isoelectric focusing | - |
5.88 |
Gluconobacter oxydans | isoelectric focusing | - |
5.9 |
Organism | Comment | Expression |
---|---|---|
Gluconobacter oxydans | induced by D-arabinitol | up |
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 |