Protein Variants | Comment | Organism |
---|---|---|
K274M | site-directed mutagenesis, the mutant enzyme shows increased activity and altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
K74M/N276D | site-directed mutagenesis, the mutant enzyme shows increased activity and altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
additional information | coenzyme specificities of the NADPH-preferring xylose reductase and the NAD+-dependent xylitol dehydrogenase, EC 1.1.1.9, are targeted in previous studies by protein design or evolution with the aim of improving the recycling of NADH or NADPH in their two-step pathway, converting xylose to xylulose. Yeast strains expressing variant pairs of both enzymes that according to in vitro kinetic data are suggested to be much better matched in coenzyme usage than the corresponding pair of wild-type enzymes, exhibit widely varying capabilities for xylose fermentation, bi-substrate kinetic analysis, and statistical analysis, overview. Engineered strains of Saccharomyces cerevisiae have engineered forms of xylose reductase or xylose dehydrogenase and improved performance in xylose fermentation | Saccharomyces cerevisiae |
N276D | site-directed mutagenesis, the mutant enzyme shows increased activity and altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of wild-type and mutant enzymes, detailed overview | Saccharomyces cerevisiae | |
0.003 | - |
NADPH | pH 7.0, 25°C, wild-type enzyme | Saccharomyces cerevisiae | |
0.017 | - |
NADPH | pH 7.0, 25°C, mutant N276D | Saccharomyces cerevisiae | |
0.026 | - |
NADH | pH 7.0, 25°C, mutant N276D | Saccharomyces cerevisiae | |
0.038 | - |
NADH | pH 7.0, 25°C, wild-type enzyme | Saccharomyces cerevisiae | |
0.041 | - |
NADH | pH 7.0, 25°C, mutant K274M/N276D | Saccharomyces cerevisiae | |
0.075 | - |
NADPH | pH 7.0, 25°C, mutant K274M | Saccharomyces cerevisiae | |
0.128 | - |
NADPH | pH 7.0, 25°C, mutant K274M/N276D | Saccharomyces cerevisiae | |
0.38 | - |
NADH | pH 7.0, 25°C, mutant K274M | Saccharomyces cerevisiae | |
96 | - |
D-xylose | pH 7.0, 25°C, wild-type enzyme, with NADPH | Saccharomyces cerevisiae | |
99 | - |
D-xylose | pH 7.0, 25°C, mutant N276D, with NADH | Saccharomyces cerevisiae | |
106 | - |
D-xylose | pH 7.0, 25°C, mutant K274M/N276D, with NADH | Saccharomyces cerevisiae | |
142 | - |
D-xylose | pH 7.0, 25°C, wild-type enzyme, with NADH | Saccharomyces cerevisiae | |
170 | - |
D-xylose | pH 7.0, 25°C, mutant N276D, with NADPH | Saccharomyces cerevisiae | |
229 | - |
D-xylose | pH 7.0, 25°C, mutant K274M, with NADH | Saccharomyces cerevisiae | |
506 | - |
D-xylose | pH 7.0, 25°C, mutant K274M, with NADPH | Saccharomyces cerevisiae | |
722 | - |
D-xylose | pH 7.0, 25°C, mutant K274M/N276D, with NADPH | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
xylitol + NAD+ | Saccharomyces cerevisiae | - |
D-xylose + NADH + H+ | - |
r | |
xylitol + NADP+ | Saccharomyces cerevisiae | - |
D-xylose + NADPH + H+ | NADPH is the preferred cofactor | r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
xylitol + NAD(P)+ = D-xylose + NAD(P)H + H+ | ordered bi-substrate mechanism in which the coenzyme binds first | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
xylitol + NAD+ | - |
Saccharomyces cerevisiae | D-xylose + NADH + H+ | - |
r | |
xylitol + NADP+ | - |
Saccharomyces cerevisiae | D-xylose + NADPH + H+ | NADPH is the preferred cofactor | r |
Synonyms | Comment | Organism |
---|---|---|
xylose reductase | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Saccharomyces cerevisiae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3 | 6 | D-xylose | pH 7.0, 25°C, mutant K274M, with NADPH | Saccharomyces cerevisiae | |
11 | - |
D-xylose | pH 7.0, 25°C, wild-type enzyme, with NADH | Saccharomyces cerevisiae | |
12 | - |
D-xylose | pH 7.0, 25°C, mutant K274M/N276D, with NADH | Saccharomyces cerevisiae | |
13 | - |
D-xylose | pH 7.0, 25°C, wild-type enzyme, with NADPH | Saccharomyces cerevisiae | |
14 | - |
D-xylose | pH 7.0, 25°C, mutant N276D, with NADH | Saccharomyces cerevisiae | |
19 | - |
D-xylose | pH 7.0, 25°C, mutant K274M, with NADH | Saccharomyces cerevisiae | |
30 | - |
D-xylose | pH 7.0, 25°C, mutant K274M/N276D, with NADPH | Saccharomyces cerevisiae | |
37 | - |
D-xylose | pH 7.0, 25°C, mutant N276D, with NADPH | Saccharomyces cerevisiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Saccharomyces cerevisiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Saccharomyces cerevisiae | |
NADH | - |
Saccharomyces cerevisiae | |
NADP+ | - |
Saccharomyces cerevisiae | |
NADPH | preferred cofactor | Saccharomyces cerevisiae |