Cloned (Comment) | Organism |
---|---|
recombinant expression of the enzyme, constructs prepared for: (a) binding of xylose reductase to cell wall glucan through the fusion with the Pir4 cell wall protein (Pir-XR) expressed under regulation by the GAL1 promoter, (b) binding of xylose reductase to cell wall glucan through the Ccw12 GPI domain (XR-GPI) expressed under regulation by the GAL1 promoter, (c) secretion of xylose reductase into the medium (XR) expressed under regulation by the GAL1 promoter, (d) binding of xylose reductase to cell wall glucan through the Ccw12 GPI domain (XR-GPI) expressed under regulation by the PHO5 promoter. Relative activity of XR-GPI under PHO5 promoter at different concentrations of phosphate in growth media, overview | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
additional information | the enzyme is labelled by the insertion of the hemagglutinin (HA) tag between the end of the secretion signal sequence and the original XR. The obtained proteinis designated as XR-GPI. The second construct is obtained by fusing Pir4/Ccw5 protein to the N-terminus of XR and the addition of the HA tag to the C-terminus of the construct. This protein is named Pir-XR | Saccharomyces cerevisiae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | - |
Saccharomyces cerevisiae | |
additional information | wild-type and engineered enzymes are resistant to the addition of EDTA and are not much impaired by non-ionic detergents like Tweens or Triton X-100, but they are sensitive to SDS, PMSF and 2-mercaptoethanol | Saccharomyces cerevisiae | |
PMSF | - |
Saccharomyces cerevisiae | |
SDS | - |
Saccharomyces cerevisiae | |
Triton X-100 | slight inhibition | Saccharomyces cerevisiae | |
Tween 20 | slight to moderate inhibition | Saccharomyces cerevisiae | |
Tween 80 | slight to moderate inhibition | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-fructose + NADPH + H+ | Saccharomyces cerevisiae | - |
sorbitol + NADP+ | - |
r | |
D-fructose + NADPH + H+ | Saccharomyces cerevisiae ATCC 204508 | - |
sorbitol + NADP+ | - |
r | |
D-galactose + NADPH + H+ | Saccharomyces cerevisiae | - |
galactitol + NADP+ | - |
r | |
D-galactose + NADPH + H+ | Saccharomyces cerevisiae ATCC 204508 | - |
galactitol + NADP+ | - |
r | |
D-glucose + NADPH + H+ | Saccharomyces cerevisiae | - |
glucitol + NADP+ | - |
r | |
D-glucose + NADPH + H+ | Saccharomyces cerevisiae ATCC 204508 | - |
glucitol + NADP+ | - |
r | |
D-xylose + NADPH + H+ | Saccharomyces cerevisiae | - |
xylitol + NADP+ | - |
r | |
D-xylose + NADPH + H+ | Saccharomyces cerevisiae ATCC 204508 | - |
xylitol + NADP+ | - |
r | |
L-arabinose + NADPH + H+ | Saccharomyces cerevisiae | - |
arabitol + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | P38715 | - |
- |
Saccharomyces cerevisiae ATCC 204508 | P38715 | - |
- |
Storage Stability | Organism |
---|---|
-20°C, purified recombinant wild-type enzyme has a half-life of 36 h, engineered enzyme Pir-XR of 84 h, and engineered enzyme XR-GPI of 72 h | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-fructose + NADPH + H+ | - |
Saccharomyces cerevisiae | sorbitol + NADP+ | - |
r | |
D-fructose + NADPH + H+ | - |
Saccharomyces cerevisiae ATCC 204508 | sorbitol + NADP+ | - |
r | |
D-galactose + NADPH + H+ | - |
Saccharomyces cerevisiae | galactitol + NADP+ | - |
r | |
D-galactose + NADPH + H+ | - |
Saccharomyces cerevisiae ATCC 204508 | galactitol + NADP+ | - |
r | |
D-glucose + NADPH + H+ | - |
Saccharomyces cerevisiae | glucitol + NADP+ | - |
r | |
D-glucose + NADPH + H+ | - |
Saccharomyces cerevisiae ATCC 204508 | glucitol + NADP+ | - |
r | |
D-xylose + NADPH + H+ | - |
Saccharomyces cerevisiae | xylitol + NADP+ | - |
r | |
D-xylose + NADPH + H+ | - |
Saccharomyces cerevisiae ATCC 204508 | xylitol + NADP+ | - |
r | |
L-arabinose + NADPH + H+ | - |
Saccharomyces cerevisiae | arabitol + NADP+ | - |
r | |
additional information | Gre3 is generally described as aldose reductase, the enzyme is not particularly specific for xylose but has significant activity with other sugars, as well | Saccharomyces cerevisiae | ? | - |
- |
|
additional information | Gre3 is generally described as aldose reductase, the enzyme is not particularly specific for xylose but has significant activity with other sugars, as well | Saccharomyces cerevisiae ATCC 204508 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
GRE3 | - |
Saccharomyces cerevisiae |
More | see also EC 1.1.1.21 | Saccharomyces cerevisiae |
xylose reductase | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
- |
Saccharomyces cerevisiae |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 70 | 50% of maximal activity at 30°C and 70°C | Saccharomyces cerevisiae |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
purified recombinant wild-type enzyme has a half-life of 60 min, engineered enzyme Pir-XR of 120 min, and engineered enzyme XR-GPI of 120 min | Saccharomyces cerevisiae |
40 | - |
purified recombinant wild-type enzyme has a half-life of 45 min, engineered enzyme Pir-XR of 100 min, and engineered enzyme XR-GPI of 100 min | Saccharomyces cerevisiae |
50 | - |
purified recombinant wild-type enzyme has a half-life of 30 min, engineered enzyme Pir-XR of 60 min, and engineered enzyme XR-GPI of 40 min | Saccharomyces cerevisiae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.06 | - |
NADPH | pH 5.0, 30°C, recombinant wild-type enzyme | Saccharomyces cerevisiae | |
0.14 | - |
NADPH | pH 5.0, 30°C, recombinant engineered XR-GPI enzyme | Saccharomyces cerevisiae | |
0.17 | - |
NADPH | pH 5.0, 30°C, recombinant engineered Pir-XR enzyme | Saccharomyces cerevisiae | |
157 | - |
D-xylose | pH 5.0, 30°C, recombinant wild-type enzyme | Saccharomyces cerevisiae | |
227 | - |
D-xylose | pH 5.0, 30°C, recombinant engineered Pir-XR enzyme | Saccharomyces cerevisiae | |
343 | - |
D-xylose | pH 5.0, 30°C, recombinant engineered XR-GPI enzyme | Saccharomyces cerevisiae | |
350 | - |
D-galactose | pH 5.0, 30°C, recombinant engineered Pir-XR enzyme | Saccharomyces cerevisiae | |
360 | - |
D-galactose | pH 5.0, 30°C, recombinant wild-type enzyme | Saccharomyces cerevisiae | |
410 | - |
D-galactose | pH 5.0, 30°C, recombinant engineered XR-GPI enzyme | Saccharomyces cerevisiae | |
420 | - |
D-fructose | pH 5.0, 30°C, recombinant engineered Pir-XR enzyme | Saccharomyces cerevisiae | |
466 | - |
D-fructose | pH 5.0, 30°C, recombinant wild-type enzyme | Saccharomyces cerevisiae | |
530 | - |
D-fructose | pH 5.0, 30°C, recombinant engineered XR-GPI enzyme | Saccharomyces cerevisiae | |
790 | - |
D-glucose | pH 5.0, 30°C, recombinant wild-type enzyme | Saccharomyces cerevisiae | |
900 | - |
L-arabinose | pH 5.0, 30°C, recombinant engineered XR-GPI enzyme | Saccharomyces cerevisiae | |
1000 | - |
L-arabinose | pH 5.0, 30°C, recombinant engineered Pir-XR enzyme | Saccharomyces cerevisiae | |
1030 | - |
L-arabinose | pH 5.0, 30°C, recombinant wild-type enzyme | Saccharomyces cerevisiae | |
1920 | - |
D-glucose | pH 5.0, 30°C, recombinant engineered Pir-XR enzyme | Saccharomyces cerevisiae | |
3190 | - |
D-glucose | pH 5.0, 30°C, recombinant engineered XR-GPI enzyme | Saccharomyces cerevisiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
sugar reduction | Saccharomyces cerevisiae |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3.8 | 6.2 | 50% of maximal activity at pH 3.8 and pH 6.2, sugar reduction | Saccharomyces cerevisiae |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
3.5 | - |
purified recombinant wild-type and engineered enzymes have a half-life of 1.2 h | Saccharomyces cerevisiae |
5 | - |
purified recombinant wild-type enzyme has a half-life of 10 h, engineered enzyme Pir-XR of 30 h, and engineered enzyme XR-GPI of 30 h | Saccharomyces cerevisiae |
6 | - |
purified recombinant wild-type enzyme has a half-life of 5 h, engineered enzyme Pir-XR of 20 h, and engineered enzyme XR-GPI of 20 h | Saccharomyces cerevisiae |
7 | - |
purified recombinant wild-type enzyme has a half-life of 4 h, engineered enzyme Pir-XR of 7.5 h, and engineered enzyme XR-GPI of 12.5 h | Saccharomyces cerevisiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Saccharomyces cerevisiae | |
NADPH | - |
Saccharomyces cerevisiae |