1.1.1.431: D-xylose reductase (NADPH)
This is an abbreviated version!
For detailed information about D-xylose reductase (NADPH), go to the full flat file.
Reaction
Synonyms
ALR1, Cb-XR, CbXR, CtXR, D-xylose reductase, D-xylose reductase 1, D-xylose reductase 2, D-xylose reductase 3, DnXR, GRE3, monospecific xylose reductase, More, msXR, NAD(P)H-dependent D-xylose reductase, NAD(P)H-dependent xylose reductase, NADPH dependent D-xylose reductase, NADPH-dependent D-xylose reductase II,III, NADPH-dependent xylose reductase, NADPH-preferring xylose reductase, NRRL3_10868, SsXR, Texr, TrxR, XR1, XR2, XR3, XRTL, XYL1, xylose reductase, xyr8, XyrA, XyrB
ECTree
1.1.1.431 D-xylose reductase (NADPH)Advanced search results
results (
results (Engineering
Engineering on EC 1.1.1.431 - D-xylose reductase (NADPH)
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
K274R
D47A
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
F111A
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
F128A
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
F221A
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
H110A
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
K21A
-
mutation reverses the cofactor specificity from major NADP- to NAD-dependent
K270N
-
mutation reverses the cofactor specificity from major NADP- to NAD-dependent
L224A
site-directed mutagenesis of the substrate binding residue, the mutant shows 35% XR activity compared to the wild-type enzyme
N306A
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
W20A
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
W311A
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
W79A
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
D47A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
F221A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
H110A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
L224A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows 35% XR activity compared to the wild-type enzyme
-
N306A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
D47A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
F221A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
H110A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
L224A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows 35% XR activity compared to the wild-type enzyme
-
N306A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
D47A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
F221A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
H110A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
L224A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows 35% XR activity compared to the wild-type enzyme
-
N306A
-
site-directed mutagenesis of the substrate binding residue, the mutant shows almost complete loss of activity
-
D50A
mutant shows 31% and 18% of the wild-type catalytic-centre activities for xylose reduction and xylitol oxidation respectively, consistent with a decrease in the rates of the chemical steps caused by the mutation, but no change in the apparent substrate binding constants and the pattern of substrate specificities
K274R/N276D
N309A
the 30fold preference of the wild-type for D-galactose compared with 2-deoxy-D-galactose is lost completely in the mutant. Replacement of Asn309 with alanine or aspartic acid disrupts the function of the original side chain in donating a hydrogen atom for bonding with the substrate C-2(R) hydroxy group, thus causing a loss of transition-state stabilization energy of 89 kJ/mol
N309D
the 30fold preference of the wild-type for D-galactose compared with 2-deoxy-D-galactose is lost completely in the mutant. Comparison of the 2.4 A X-ray crystal structure of mutant N309D bound to NAD+ with the previous structure of the wild-type holoenzyme reveals no major structural perturbations. Replacement of Asn309 with alanine or aspartic acid disrupts the function of the original side chain in donating a hydrogen atom for bonding with the substrate C-2(R) hydroxy group, thus causing a loss of transition-state stabilization energy of 89 kJ/mol
W23F
mutant catalyses NADH-dependent reduction of xylose with 4% of the wild-type efficiency (kcat/Km), but improves the wild-type selectivity for utilization of ketones, relative to xylose, by factors of 156
W23Y
mutant catalyses NADH-dependent reduction of xylose with 1% of the wild-type efficiency (kcat/Km), but improves the wild-type selectivity for utilization of ketones, relative to xylose, by factors of 471
additional information
K274R
-
mutation introduced to change the specificity toward NADH. Fermentation with the mutant strain shows a 2.8fold reduction in xylitol accumulation and 4.5fold increase in citric acid production compared to the wild-type strain
K274R
-
mutation introduced to change the specificity toward NADH. Fermentation with the mutant strain shows a 2.8fold reduction in xylitol accumulation and 4.5fold increase in citric acid production compared to the wild-type strain
-
K274R/N276D
-
structure-guided site-directed mutagenesis, change of the coenzyme preference of the xyluose reductase about 170fold from NADPH in the wild-type to NADH, which, in spite of the structural modifications introduced, has retained the original catalytic efficiency for reduction of xylose by NADH
K274R/N276D
NADH-specific mutant, Saccharomyces cerevisiae expressing mutant K274R/N276D exhibits intracellular activities of 0.94 U/mg 1.07 U/mg with NADPH and NADH, respectively
additional information
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production of xylitol from D-xylose and D-glucose with recombinant Corynebacterium glutamicum, the strain is engineered to express the xylose reductase gene XYL1of Pichia stipitis, and produces xylose reductase with a specific activity of ca. 0.6 U/mg protein. Due to the absence of xylose isomerase and xylitol dehydrogenase genes, loose catabolite repression, high NADPH regeneration capacity, and tolerance against sugar-induced osmotic stress, the recombinant biocatalyst is able to efficiently produce xylitol from D-xylose using glucose as source of reducing equivalents
additional information
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production of xylitol from D-xylose and D-glucose with recombinant Corynebacterium glutamicum, the strain is engineered to express the xylose reductase gene XYL1of Pichia stipitis, and produces xylose reductase with a specific activity of ca. 0.6 U/mg protein. Due to the absence of xylose isomerase and xylitol dehydrogenase genes, loose catabolite repression, high NADPH regeneration capacity, and tolerance against sugar-induced osmotic stress, the recombinant biocatalyst is able to efficiently produce xylitol from D-xylose using glucose as source of reducing equivalents
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additional information
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the native xylose reductase gene Kmxyl1 of the Kluyveromyces marxianus strain YHJ010 is substituted with xylose reductase or its mutants N272D, K21A/N272D, or K270M from Pichia stipitis, i.e. Scheffersomyces stipitis, resulting in Kluyveromyces marxianus strains YZB013, YZB014, and YZB015. The ability of the resultant recombinant strains to assimilate xylose to produce xylitol and ethanol at elevated temperature is greatly improved, overview. But the strain YZB015 expressing a mutant PsXR K21A/N272D, with which co-enzyme preference is completely reversed from NADPH to NADH, fails to ferment due to the low expression
additional information
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the native xylose reductase gene Kmxyl1 of the Kluyveromyces marxianus strain YHJ010 is substituted with xylose reductase or its mutants N272D, K21A/N272D, or K270M from Pichia stipitis, i.e. Scheffersomyces stipitis, resulting in Kluyveromyces marxianus strains YZB013, YZB014, and YZB015. The ability of the resultant recombinant strains to assimilate xylose to produce xylitol and ethanol at elevated temperature is greatly improved, overview. But the strain YZB015 expressing a mutant PsXR K21A/N272D, with which co-enzyme preference is completely reversed from NADPH to NADH, fails to ferment due to the low expression
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additional information
efficient biosynthesis of xylitol from xylose by coexpression of xylose reductase (XR) from Rhizopus oryzae and glucose dehydrogenase (GDH) from Exiguobacterium sibiricum, the latter is used for cofactor regeneration, in Escherichia coli strain BL21(DE3)/pCDFDuet-1-XR-GDH, from Escherichia coli strains BL21(DE3)/pET28b(+)-XR and BL21(DE3)/pET28b(+)-GDH. The xylitol yield of the coupled system is maximal at pH 8.0 and 30°C, and at a unit ratio of GDH to XR concentration of 4:1, indicating that the regeneration of coenzyme had a significant effect on xylitol synthesis. Method optimization, overview
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
additional information
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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
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additional information
construction of a set of recombinant Saccharomyces cerevisiae carrying a mutated strictly NADPH-dependent xylose reductase and NADP+-dependent xylitol dehydrogenase genes with overexpression of endogenous xylulokinase (XK), effects of complete NADPH/NADP+ recycling on ethanol fermentation and xylitol accumulation, overview. The mutated strains demonstrate 0% and 10% improvement in ethanol production, and reduced xylitol accumulation, ranging 34.4-54.7% compared with the control strain
additional information
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construction of a set of recombinant Saccharomyces cerevisiae carrying a mutated strictly NADPH-dependent xylose reductase and NADP+-dependent xylitol dehydrogenase genes with overexpression of endogenous xylulokinase (XK), effects of complete NADPH/NADP+ recycling on ethanol fermentation and xylitol accumulation, overview. The mutated strains demonstrate 0% and 10% improvement in ethanol production, and reduced xylitol accumulation, ranging 34.4-54.7% compared with the control strain
additional information
enzyme XRTL can therefore be used in a cell-free xylitol production process or as part of a pathway for utilization of xylose from lignocellulosic waste. Ferulic acid is an inhibitor of the lignocellulosic activity
additional information
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enzyme XRTL can therefore be used in a cell-free xylitol production process or as part of a pathway for utilization of xylose from lignocellulosic waste. Ferulic acid is an inhibitor of the lignocellulosic activity
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additional information
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the mutant Candida tenuis enzyme is modified in its cofactor specificity showing preference for NADPH compared to NADH in the D-xylose reduction reaction, genetic metabolic engineering for improvement of xylose metabolism and fermentation in wild-type Saccharomyces cerevisiae strains, which are not able to naturally metabolize D-xylulose, overview
additional information
efficient uptake and reduction of xylose photoautotrophically in Synechococcus elongatus strain PCC7942 are demonstrated upon introduction of an effective xylose transporter from Escherichia coli (Ec-XylE) and the NADPH-dependent xylose reductase from Candida boidinii (Cb-XR). Simultaneous activation of xylose uptake and matching of cofactor specificity enables an average xylitol yield of 0.9 g/g xylose and a maximum productivity of about 0.15 g/l/day/OD with increased level of xylose supply. High-density conversion of xylose to xylitol using concentrated resting cells further pushes the titer of xylitol formation to 33 g/l in six days with 85% of maximum theoretical yield. Comparison of the efficiencies of the two routes for xylitol biosynthesis, detailed overview
additional information
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efficient uptake and reduction of xylose photoautotrophically in Synechococcus elongatus strain PCC7942 are demonstrated upon introduction of an effective xylose transporter from Escherichia coli (Ec-XylE) and the NADPH-dependent xylose reductase from Candida boidinii (Cb-XR). Simultaneous activation of xylose uptake and matching of cofactor specificity enables an average xylitol yield of 0.9 g/g xylose and a maximum productivity of about 0.15 g/l/day/OD with increased level of xylose supply. High-density conversion of xylose to xylitol using concentrated resting cells further pushes the titer of xylitol formation to 33 g/l in six days with 85% of maximum theoretical yield. Comparison of the efficiencies of the two routes for xylitol biosynthesis, detailed overview
