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Enzyme Nomenclature
Information on EC 1.1.1.307 - D-xylose reductase
Word Map on EC 1.1.1.307
- 1.1.1.307
- ethanol
- l-arabinose
-
nadph-dependent
-
pichia
- synthesis
- reesei
- stipitis
- trichoderma
- l-arabitol
- cellulase
- galactitol
-
pachysolen
-
kluyveromyces
-
oxygen-limited
-
hypocrea
- tannophilus
-
nadph-linked
- d-galactose
- jecorina
- pentitols
- cellobiohydrolases
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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
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EC NUMBER 
COMMENTARY 
1.1.1.307
-
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RECOMMENDED NAME
GeneOntology No.
D-xylose reductase
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
xylitol + NAD(P)+ = D-xylose + NAD(P)H + H+
-
-
-
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xylitol + NAD(P)+ = D-xylose + NAD(P)H + H+
enzymic mechanism in which a catalytic proton bridge from the protonated side chain of Lys80 to the carbonyl group adjacent to the hydride acceptor carbonyl facilitates the chemical reaction step. His113 contributes to positioning of the 9,10-phenanthrenequinone substrate for catalysis. Tyr51 controls release of the hydroquinone product. The proposed chemistry involves delivery of both hydrogens required for reduction of the alpha-dicarbonyl substrate to the carbonyl group undergoing stereoselective transformation. Hydride transfer from NADH probably precedes the transfer of a proton from Tyr51
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xylitol + NAD(P)+ = D-xylose + NAD(P)H + H+
ordered bi-substrate mechanism in which the coenzyme binds first
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xylitol + NAD+ = D-xylose + NADH + H+
chemical mechanism of carbonyl reduction by xylose reductase in which transfer of hydride ion is a partially rate-limiting step and precedes the proton-transfer step
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xylitol + NAD+ = D-xylose + NADH + H+
-
-
-
-
xylitol + NADP+ = D-xylose + NADPH + H+
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-
-
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xylitol + NADP+ = D-xylose + NADPH + H+
kinetic mechanism of xylose reductase is iso-ordered bi bi
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
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SYSTEMATIC NAME
IUBMB Comments
xylitol:NAD(P)+ oxidoreductase
Xylose reductase catalyses the initial reaction in the xylose utilization pathway, the NAD(P)H dependent reduction of xylose to xylitol.
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
ssp. mobilis, and strain A3, an engineered strain ZM4 that is adapted to 5% D-xylose, gene ZMO0976
UniProt
ssp. mobilis, and strain A3, an engineered strain ZM4 that is adapted to 5% D-xylose, gene ZMO0976
UniProt
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
metabolism
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
metabolism
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
metabolism
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
metabolism
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
metabolism
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
metabolism
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
metabolism
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
metabolism
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
metabolism
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
metabolism
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
metabolism
Torulopsis molishiama
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
metabolism
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key enzymes for xylitol production in yeasts are xylose reductase and xylitol dehydrogenase, EC 1.1.1.9, overview
metabolism
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xylose reductase is the first enzyme in D-xylose metabolism, catalyzing the reduction of D-xylose to xylitol
metabolism
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
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metabolism
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
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metabolism
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xylose reductase is the first enzyme in D-xylose metabolism, catalyzing the reduction of D-xylose to xylitol
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metabolism
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
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metabolism
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
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metabolism
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
metabolism
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
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metabolism
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
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metabolism
Torulopsis molishiama 55
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xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2-deoxy-D-galactose + NADH + H+
?
-
dual specific xylose reductase (dsXR)
-
-
?
2-deoxy-D-galactose + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
2-deoxy-D-glucose + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
2-deoxy-D-ribose + NADH + H+
?
-
dual specific xylose reductase (dsXR)
-
-
?
2-deoxy-D-ribose + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
butanal + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-erythrose + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-erythrose + NADPH + H+
erythritol + NADP+
-
catalytic efficiency is 100fold higher than the catalytic efficiency for D-xylose
-
-
?
D-fucose + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-glucosone + NADPH + H+
D-fructose + NADP+
-
catalytic efficiency is 22fold higher than the catalytic efficiency for D-xylose
-
-
?
D-lyxose + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-mannose + NADH + H+
?
8% of the activity compared to D-xylose (with NADH as cofactor)
-
-
?
L-arabinose + NADH + H+
arabinitol + NAD+
-
low activity in direction of arabinitol oxidation. At pH 6.0 polyol oxidation is not observed, but between pH 8 and 9 the enzyme oxidizes the polyol
-
-
r
L-arabinose + NADH + H+
L-arabinitol + NAD+
-
dual specific xylose reductase (dsXR)
-
-
?
L-arabinose + NADPH + H+
arabinitol + NADP+
-
low activity in direction of arabinitol oxidation. At pH 6.0 polyol oxidation is not observed, but between pH 8 and 9 the enzyme oxidizes the polyol
-
-
r
L-lyxose + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
methylglyoxal + NADPH + H+
?
-
catalytic efficiency is 20fold higher than the catalytic efficiency for D-xylose
-
-
?
pentanal + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
phenylglyoxal + NADPH + H+
?
-
catalytic efficiency is 17fold higher than the catalytic efficiency for D-xylose
-
-
?
propionaldehyde + NADH + H+
?
-
dual specific xylose reductase (dsXR)
-
-
?
propionaldehyde + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
pyridine-2-aldehyde + NADPH + H+
?
-
catalytic efficiency is 7fold higher than the catalytic efficiency for D-xylose
-
-
?
valeraldehyde + NADPH + H+
?
-
catalytic efficiency is 13fold higher than the catalytic efficiency for D-xylose
-
-
?
xylosone + NADPH + H+
?
-
catalytic efficiency is 20fold higher than the catalytic efficiency for D-xylose
-
-
?
5-hydroxymethylfurfural + NADH + H+
(furan-2,5-diyl)dimethanol + NAD+
low activity
-
-
r
5-hydroxymethylfurfural + NADH + H+
(furan-2,5-diyl)dimethanol + NAD+
low activity
-
-
r
benzaldehyde + NADH + H+
benzyl alcohol + NAD+
best substrate
-
-
r
benzaldehyde + NADH + H+
benzyl alcohol + NAD+
best substrate
-
-
r
D-arabinose + NADPH + H+
?
about 60% of the activity compared to D-xylose
-
-
?
D-arabinose + NADPH + H+
?
about 60% of the activity compared to D-xylose
-
-
?
D-galactose + NADH + H+
?
48% of the activity compared to D-xylose (with NADH as cofactor)
-
-
?
D-galactose + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-galactose + NADPH + H+
?
about 70% of the activity compared to D-xylose
-
-
?
D-galactose + NADPH + H+
?
-
D-xylose reductase 1: 49% of the activity with D-xylose, D-xylose reductase 2: 40% of the activity with D-xylose, D-xylose reductase 3: 33% of the activity with D-xylose
-
-
?
D-galactose + NADPH + H+
?
-
D-xylose reductase 1: 49% of the activity with D-xylose, D-xylose reductase 2: 40% of the activity with D-xylose, D-xylose reductase 3: 33% of the activity with D-xylose
-
-
?
D-galactose + NADPH + H+
?
-
catalytic efficiency is 9.1% of the catalytic efficiency for D-xylose
-
-
?
D-glucose + NADH + H+
?
10 of the activity compared to D-xylose (with NADH as cofactor)
-
-
?
D-glucose + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-glucose + NADPH + H+
?
about 15% of the activity compared to D-xylose
-
-
?
D-glucose + NADPH + H+
?
-
D-xylose reductase 1: 10% of the activity with D-xylose, D-xylose reductase 2: 11% of the activity with D-xylose, D-xylose reductase 3: 11% of the activity with D-xylose
-
-
?
D-glucose + NADPH + H+
?
-
D-xylose reductase 1: 10% of the activity with D-xylose, D-xylose reductase 2: 11% of the activity with D-xylose, D-xylose reductase 3: 11% of the activity with D-xylose
-
-
?
D-glucose + NADPH + H+
?
-
catalytic efficiency is 3.3% of the catalytic efficiency for D-xylose
-
-
?
D-ribose + NADPH + H+
?
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-ribose + NADPH + H+
?
about 90% of the activity compared to D-xylose
-
-
?
D-ribose + NADPH + H+
?
-
catalytic efficiency is 41% of the catalytic efficiency for D-xylose
-
-
?
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
-
xylose reductase, using either NADH or NADPH, reduces D-xylose to xylitol, subsequently xylitol is oxidized to D-xylulose by a NAD+-linked xylulose dehydrogenase, EC 1.1.1.9
-
-
?
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
-
xylose reductase, using either NADH or NADPH, reduces D-xylose to xylitol, subsequently xylitol is oxidized to D-xylulose by a NAD+-linked xylulose dehydrogenase, EC 1.1.1.9
-
-
?
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
-
xylose reductase, using either NADH or NADPH, reduces D-xylose to xylitol, subsequently xylitol is oxidized to D-xylulose by a NAD+-linked xylulose dehydrogenase, EC 1.1.1.9
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
-
using a modified iterative protein redesign and optimization workflow, a sets of mutations is identified that change the nicotinamide cofactor specificity of xylose reductase (CbXR) from its physiological preference for NADPH, to the alternate cofactor NADH
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
-
reaction is catalyzed by dual specific xylose reductase (dsXR), reaction is not catalyzed by NADPH-dependent monospecific xylose reductase (msXR)
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
the enzyme specifically transfers the 4-pro-R hydrogen from the C-4 of the nicotinamide ring to the re face of the carbonyl carbon of the substrate
-
-
r
D-xylose + NADH + H+
xylitol + NAD+
the enzyme specifically transfers the 4-pro-R hydrogen from the C-4 of the nicotinamide ring to the re face of the carbonyl carbon of the substrate
-
-
r
D-xylose + NADH + H+
xylitol + NAD+
-
dual (NADH and NADPH) coenzyme specificity
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
kcat of wilde-type enzyme increases by a factor of 1.73 when NADPH replaces NADH
-
-
r
D-xylose + NADH + H+
xylitol + NAD+
dual coenzyme specificity, Km for NADPH: 0.0455 mM, Km for NADH: 0.162 mM
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
dual coenzyme specificity, Km for NADPH: 0.0455 mM, Km for NADH: 0.162 mM
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
-
catalytic efficiency for NADPH is more than 100fold higher than the catalytic efficiency for NADH
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
wild-type TeXR shows dual coenzyme specificity but is preferentially NADPH-dependent, with affinity for NADPH being 1.1fold higher than NADH and catalytic efficiency (kcat/Km) 24.5fold higher with NADPH as coenzyme. Affinity for xylose is 3.6fold higher with NADPH as coenzyme
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
NADPH is the preferred cofactor
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
-
active with both NADPH and NADH as coenzyme. The activity with NADH is approximately 70% of that with NADPH for the various aldose substrates. Rate of xylitol oxidation is 4% of the rate of D-xylose reduction. At pH 6.0 polyol oxidation is not observed, but between pH 8 and 9 the enzyme oxidizes the polyol
-
-
r
D-xylose + NADH + H+
xylitol + NAD+
-
wild-type enzyme prefers NADPH over NADH
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
using a modified iterative protein redesign and optimization workflow, a sets of mutations is identified that change the nicotinamide cofactor specificity of xylose reductase (CbXR) from its physiological preference for NADPH, to the alternate cofactor NADH
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
reaction is catalyzed by NADPH-dependent monospecific xylose reductase (msXR), and by dual specific xylose reductase (dsXR)
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
xylose reductases catalyse the initial reaction in the xylose utilisation pathway, the NAD(P)H dependent reduction of xylose to xylitol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
catalytic efficiency (kcat/Km) in D-xylose reduction at pH 7 is more than 60fold higher than that in xylitol oxidation. The enzyme prefers NADPH approximately 2fold to NADH
-
-
r
D-xylose + NADPH + H+
xylitol + NADP+
-
dual (NADH and NADPH) coenzyme specificity
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
kcat of wild-type enzyme increases by a factor of 1.73 when NADPH replaces NADH
-
-
r
D-xylose + NADPH + H+
xylitol + NADP+
xylose reductases catalyse the initial reaction in the xylose utilisation pathway, the NAD(P)H dependent reduction of xylose to xylitol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
key enzyme in xylose metabolism
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
dual coenzyme specificity, Km for NADPH: 0.0455 mM, Km for NADH: 0.162 mM
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
key enzyme in xylose metabolism
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
dual coenzyme specificity, Km for NADPH: 0.0455 mM, Km for NADH: 0.162 mM
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
catalytic efficiency for NADPH is more than 100fold higher than the catalytic efficiency for NADH
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
kinetic mechanism of xylose reductase is iso-ordered bi bi
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
expression of Texr is inducible by the same carbon sources responsible for the induction of genes encoding enzymes relevant to lignocellulose hydrolysis, suggesting a coordinated expression of intracellular and extracellular enzymes relevant to hydrolysis and metabolism of pentose sugars in Talaromyces emersonii in adaptation to its natural habitat. This indicates a potential advantage in survival and response to a nutrient-poor environment
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
wild-type TeXR shows dual coenzyme specificity but is preferentially NADPH-dependent, with affinity for NADPH being 1.1fold higher than NADH and catalytic efficiency (kcat/Km) 24.5fold higher with NADPH as coenzyme. Affinity for xylose is 3.6fold higher with NADPH as coenzyme
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
NADPH is the preferred cofactor, specific for D-xylose
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
whereas in most bacteria metabolism of D-xylose proceeds via direct isomerization to D-xylulose, catalysed by xylose isomerase (EC 5.3.1.5), in yeasts this conversion is catalysed by the sequential action of two oxidoreductases: xylose reductase and xylitol dehydrogenase (EC 1.1.1.9)
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase is one of the key enzymes for xylose fermentation
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
active with both NADPH and NADH as coenzyme. The activity with NADH is approximately 70% of that with NADPH for the various aldose substrates. Rate of xylitol oxidation is 5% of the rate of D-xylose reduction. At pH 6.0 polyol oxidation is not observed, but between pH 8 and 9 the enzyme oxidizes the polyol
-
-
r
D-xylose + NADPH + H+
xylitol + NADP+
-
wild-type enzyme prefers NADPH over NADH
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
Torulopsis molishiama
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
Torulopsis molishiama 55
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH. Pachysolen tannophilus strains, whose xylose reductases are almost equally specific for NADH and NADPH, produce xylitol and ethanol in comparable amounts, while Candida shehatae and Pichia stipitis, whose xylose reductases are more specific for NADH than for NADPH, produce predominantly ethanol
-
-
?
DL-glyceraldehyde + NADH + H+
glycerol + NAD+
-
dual specific xylose reductase (dsXR)
-
-
?
DL-glyceraldehyde + NADH + H+
glycerol + NAD+
-
low activity in direction of glycerol oxidation. At pH 6.0 polyol oxidation is not observed, but between pH 8 and 9 the enzyme oxidizes the polyol
-
-
r
DL-glyceraldehyde + NADPH + H+
glycerol + NADP+
-
NADPH-dependent monospecific xylose reductase
-
-
?
DL-glyceraldehyde + NADPH + H+
glycerol + NADP+
-
catalytic efficiency is 37fold higher than the catalytic efficiency for D-xylose
-
-
?
DL-glyceraldehyde + NADPH + H+
glycerol + NADP+
-
D-xylose reductase 1: 200% of the activity with D-xylose, D-xylose reductase 2: 268% of the activity with D-xylose, D-xylose reductase 3: 143% of the activity with D-xylose
-
-
?
DL-glyceraldehyde + NADPH + H+
glycerol + NADP+
-
D-xylose reductase 1: 200% of the activity with D-xylose, D-xylose reductase 2: 268% of the activity with D-xylose, D-xylose reductase 3: 143% of the activity with D-xylose
-
-
?
DL-glyceraldehyde + NADPH + H+
glycerol + NADP+
-
low activity in direction of glycerol oxidation. At pH 6.0 polyol oxidation is not observed, but between pH 8 and 9 the enzyme oxidizes the polyol
-
-
r
furfural + NADH + H+
(furan-2-yl)methanol + NAD+
-
-
-
r
furfural + NADH + H+
(furan-2-yl)methanol + NAD+
-
-
-
r
L-arabinose + NADPH + H+
L-arabinitol + NADP+
-
NADPH-dependent monospecific xylose reductase
-
-
?
L-arabinose + NADPH + H+
L-arabinitol + NADP+
-
catalytic efficiency is 2fold higher than the catalytic efficiency for D-xylose
-
-
?
L-arabinose + NADPH + H+
L-arabinitol + NADP+
-
D-xylose reductase 1: 117% of the activity with D-xylose, D-xylose reductase 2: 120% of the activity with D-xylose, D-xylose reductase 3: 101% of the activity with D-xylose
-
-
?
L-arabinose + NADPH + H+
L-arabinitol + NADP+
-
D-xylose reductase 1: 117% of the activity with D-xylose, D-xylose reductase 2: 120% of the activity with D-xylose, D-xylose reductase 3: 101% of the activity with D-xylose
-
-
?
L-arabinose + NADPH + H+
L-arabinitol + NADP+
-
catalytic efficiency is 41% of the catalytic efficiency for D-xylose
-
-
?
xylitol + NADP+
D-xylose + NADPH + H+
-
-
NADPH is the preferred cofactor
-
r
additional information
?
-
-
Candida intermedia produces two isoforms of xylose reductase: one is NADPH-dependent (monospecific xylose reductase, msXR), and another prefers NADH about 4fold over NADPH (dual specific xylose reductase, dsXR)
-
-
-
additional information
?
-
in vitro the enzyme also catalyzes the reduction of ketones
-
-
-
additional information
?
-
-
prefers glyceraldehyde, D-erythrose and even some aliphatic and aromatic aldehydes to the pentose sugars D-xylose and L-arabinose. Aldosones such as D-glucosone or D-xylosone are good substrates, whereas the corresponding 2-deoxy-aldose sugars are reduced at hardly detectable rates
-
-
-
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
-
xylose reductase, using either NADH or NADPH, reduces D-xylose to xylitol, subsequently xylitol is oxidized to D-xylulose by a NAD+-linked xylulose dehydrogenase, EC 1.1.1.9
-
-
?
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
-
xylose reductase, using either NADH or NADPH, reduces D-xylose to xylitol, subsequently xylitol is oxidized to D-xylulose by a NAD+-linked xylulose dehydrogenase, EC 1.1.1.9
-
-
?
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
-
xylose reductase, using either NADH or NADPH, reduces D-xylose to xylitol, subsequently xylitol is oxidized to D-xylulose by a NAD+-linked xylulose dehydrogenase, EC 1.1.1.9
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
O74237
xylose reductases catalyse the initial reaction in the xylose utilisation pathway, the NAD(P)H dependent reduction of xylose to xylitol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
O74237
xylose reductases catalyse the initial reaction in the xylose utilisation pathway, the NAD(P)H dependent reduction of xylose to xylitol
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
A9QVV8
key enzyme in xylose metabolism
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
A9QVV8
key enzyme in xylose metabolism
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
C5J3R6
expression of Texr is inducible by the same carbon sources responsible for the induction of genes encoding enzymes relevant to lignocellulose hydrolysis, suggesting a coordinated expression of intracellular and extracellular enzymes relevant to hydrolysis and metabolism of pentose sugars in Talaromyces emersonii in adaptation to its natural habitat. This indicates a potential advantage in survival and response to a nutrient-poor environment
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
whereas in most bacteria metabolism of D-xylose proceeds via direct isomerization to D-xylulose, catalysed by xylose isomerase (EC 5.3.1.5), in yeasts this conversion is catalysed by the sequential action of two oxidoreductases: xylose reductase and xylitol dehydrogenase (EC 1.1.1.9)
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
-
xylose reductase is one of the key enzymes for xylose fermentation
-
-
?
xylitol + NADP+
D-xylose + NADPH + H+
-
-
NADPH is the preferred cofactor
-
r
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
Kluyveromyces marxianus strains expressing Pichia stipitis Psxyl1 genes show reversed cofactor specificity, overview
-
NAD(P)H
-
dependent on, the wild-type enzyme prefers NADH, while a modified mutant enzyme prefers NADPH in the D-xylose reduction reaction
NADH
-
prefers NADPH approximately 2fold to NADH, largely due to better apparent binding of the phosphorylated form of the coenzyme
NADH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADH
-
catalytic efficiency for NADPH is more than 100fold higher than the catalytic efficiency for NADH
NADH
-
active with both NADPH and NADH as coenzyme. The activity with NADH is approximately 70% of that with NADPH for the various aldose substrates. The ratio of activities with NADH and NADPH is approximately constant between pH 5 and 8
NADH
-
kcat of wild-type enzyme increases by a factor of 1.73 when NADPH replaces NADH. kcat for NADPH-dependent reduction of xylose by the mutant D50A is three times that for the corresponding NADH-dependent reaction
NADH
-
transient-state and steady-state kinetic studies of the mechanism of NADH-dependent aldehyde reduction
NADH
-
wild-type enzyme prefers NADPH as cofactor. K270M mutation results in a significant increase in the Km values for both NADPH and NADH. K270R mutation increases the Km value for NADPH 25fold, while the Km for NADH only increases two-fold
NADH
-
dual specific xylose reductase (dsXR) has an about 4fold higher specificity for NADH than NADPH
NADH
catalytic efficiency is 24.5fold higher with NADPH as coenzyme than with NADH
NADH
dual coenzyme specificity, Km for NADPH: 0.0455 mM, Km for NADH: 0.162 mM
NADH
-
wild-type enzyme prefers NADPH over NADH. Mutant enzyme K270S/N272P/S271G/R276F shows a 25fold preference toward NADH over NADPH by a factor of about 13fold, or an improvement of about 42fold, as measured by the ratio of the specificity constant kcat/Km coenzyme
NADH
-
using a modified iterative protein redesign and optimization workflow, a sets of mutations is identified that change the nicotinamide cofactor specificity of xylose reductase (CbXR) from its physiological preference for NADPH, to the alternate cofactor NADH
NADPH
-
prefers NADPH approximately 2fold to NADH, largely due to better apparent binding of the phosphorylated form of the coenzyme
NADPH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADPH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADPH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADPH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADPH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADPH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADPH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADPH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADPH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADPH
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADPH
Torulopsis molishiama
-
xylose reductase in the xylitol-producing species Candida didensiae, Candida intermediae, Candida parapsilosis, Candida silvanorum, Candida tropicalis, Kluyveromyces fragilis, Kluyveromyces marxianus, Pichia guillermondii, and Torulopsis molishiama is specific for NADPH. Xylose reductase in the ethanol-producing species Pichia stipitis, Candida shehatae, and Pachysolen tannophilus is specific for both NADPH and NADH
NADPH
-
only NADPH-dependent xylose reductase is obtained under the cultivation conditions
NADPH
-
catalytic efficiency for NADPH is more than 100fold higher than the catalytic efficiency for NADH
NADPH
-
active with both NADPH and NADH as coenzyme. The activity with NADH is approximately 70% of that with NADPH for the various aldose substrates. The ratio of activities with NADH and NADPH is approximately constant between pH 5 and 8
NADPH
-
kcat of wild-type enzyme increases by a factor of 1.73 when NADPH replaces NADH. kcat for NADPH-dependent reduction of xylose by the mutant D50A is three times that for the corresponding NADH-dependent reaction
NADPH
-
wild-type enzyme prefers NADPH as cofactor. K270M mutation results in a significant increase in the Km values for both NADPH and NADH. K270R mutation increases the Km value for NADPH 25fold, while the Km for NADH only increases two-fold
NADPH
-
dual specific xylose reductase (dsXR) has an about 4fold higher specificity for NADH than NADPH. NADPH-dependent monospecific xylose reductase (msXR) shows non activity with NADH
NADPH
catalytic efficiency is 24.5fold higher with NADPH as coenzyme than with NADH. Affinity for xylose is 3.6fold higher with NADPH as coenzyme; wild-type TeXR shows dual coenzyme specificity but is preferentially NADPH-dependent, with affinity for NADPH being 1.1fold higher than NADH and catalytic efficiency (kcat/Km) 24.5fold higher with NADPH as coenzyme. Affinity for xylose is 3.6fold higher with NADPH as coenzyme. K271R/N273D double mutant displays an altered coenzyme preference with a 16fold improvement in NADH utilization relative to the wild type
NADPH
dual coenzyme specificity, Km for NADPH: 0.0455 mM, Km for NADH: 0.162 mM
NADPH
-
wild-type enzyme prefers NADPH over NADH. Mutant enzyme K270S/N272P/S271G/R276F shows a 25fold preference toward NADH over NADPH by a factor of about 13fold, or an improvement of about 42fold, as measured by the ratio of the specificity constant kcat/Km coenzyme
NADPH
-
using a modified iterative protein redesign and optimization workflow, a sets of mutations is identified that change the nicotinamide cofactor specificity of xylose reductase (CbXR) from its physiological preference for NADPH, to the alternate cofactor NADH
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Li+
Ca2+, Li+, Mg2+, Mn2+ and NH4+ at 10 mM decrease activity by 10-50%
Mg2+
Ca2+, Li+, Mg2+, Mn2+ and NH4+ at 10 mM decrease activity by 10-50%
Mn2+
Ca2+, Li+, Mg2+, Mn2+ and NH4+ at 10 mM decrease activity by 10-50%
NH4+
Ca2+, Li+, Mg2+, Mn2+ and NH4+ at 10 mM decrease activity by 10-50%
additional information
additional information
neither inhibited nor activated by EDTA at concentrations ranging from 1 to 10 mM
additional information
-
no effect: Na+, K+, NH4+, Mg2+, Ca2+ and Co2+ in the form of the chloride salt in 50 mM Tris, pH 7.0, as well the anions PO43-, SO32-, NO3-, CO32-, citrate and tetraborate in the form of the sodium salt in 50 mM phosphate buffer, pH 7.0
additional information
no increase in activity in presence of 1 mM NaCl and 1 mM MgSO4
additional information
-
no requirement for divalent cation is observed
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NADPH
-
for monospecific xylose reductase (msXR) and dual specific xylose reductase (dsXR) NADPH behaves as a competitive inhibitor against NADP+. Competitive inhibition of is observed both at unsaturating and saturating concentrations of xylitol
pyridoxal 5'-phosphate
-
gradual inactivation. NADH, ATP or 2'-AMP protects. No protection by D-xylose
NAD+
-
strong inhibition with the NADH-linked reaction, no inhibition of NADPH-linked reaction. NAD+ is a non-competitive inhibitor with respect to xylose and a competitive inhibitor with respect to NADH
NADP+
-
for monospecific xylose reductase (msXR) and dual specific xylose reductase (dsXR) NADP+ behaves as a competitive inhibitor against NADPH. Competitive inhibition of is observed both at unsaturating and saturating concentrations of xylose
NADP+
-
2 mM completely abolishes D-xylose reduction. Potent competitive inhibitor, inhibits both the NADH-dependent and the NADPH-dependent activity
NADP+
-
potent inhibitor of both the NADPH- and NADH-linked xylose reduction. Competition with NADPH and non-competitive inhibition with xylose in the NADPH-linked xylose reduction
additional information
-
no inhibition by NAD+. No effect: Na+, K+, NH4+, Mg2+, Ca2+ and Co2+ in the form of the chloride salt in 50 mM Tris, pH 7.0, as well the anions Cl-, PO43-, SO32-, NO3-, CO32-, citrate and tetraborate in the form of the sodium salt in 50 mM phosphate buffer, pH 7.0. Glucose 6-phosphate, fructose 6-phosphate, fructose 1,6-bisphosphate, 6-phosphogluconate, phosphoenolpyruvate, oxaloacetate (5 mM each) have no effect
-
additional information
-
no effect by 5 mM EDTA, 500 mM sulfate, 5 mM 2-mercaptoethanol
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
neither inhibited nor activated by EDTA at concentrations ranging from 1 to 10 mM
-
bovine serum albumin
1 mg/ml. the specific activity is increased by 20-30%
-
Triton X-100
0.1%, the specific activity is increased by 20–30%
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.058
2-deoxy-D-glucose
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.038
2-deoxy-D-ribose
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
1.8
benzaldehyde
pH 7.2, temperature not specified in the publication
4.2
furfural
pH 7.2, temperature not specified in the publication
0.126
2-deoxy-D-galactose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.221
2-deoxy-D-galactose
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
2 - 3
Butanal
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.02
D-erythrose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.033
D-erythrose
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.007
D-fucose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.01
D-fucose
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.015
D-galactose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.061
D-galactose
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.006
D-glucose
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.033
D-glucose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.068
D-ribose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.091
D-ribose
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.01
D-xylose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.016
D-xylose
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
24.56
D-xylose
pH 6.5, 37°C, cosubstrate: NADPH, wild-type enzyme
67
D-xylose
mutant K21A/N272D, cosubstrate NADH, pH 6.5, 35°C; wild-type, cosubstrate NADH, pH 6.5, 35°C
76.06
D-xylose
pH 6.5, 37°C, cosubstrate: NADH, mutant enzyme K271R/N273D
76.5
D-xylose
mutant K271R/N273, cosubstrate NADPH, pH 6.5, 37°C; pH 6.5, 37°C, cosubstrate: NADPH, mutant enzyme K271R/N273D
89.4
D-xylose
pH 6.5, 37°C, cosubstrate: NADH, wild-type enzyme; wild-type, cosubstrate NADH, pH 6.5, 37°C
168
D-xylose
-
pH 6.0, cofactor: NADPH, mutant enzyme K270S/N272P/S271G/R276F
258
D-xylose
pH 7.2, temperature not specified in the publication
291
D-xylose
-
pH 6.0, cofactor: NADH, mutant enzyme K270S/N272P/S271G/R276F
1.14
DL-glyceraldehyde
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
2.43
DL-glyceraldehyde
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.02
L-arabinose
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.028
L-arabinose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.117
L-Lyxose
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.144
L-Lyxose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.263
NADH
pH 6.5, 37°C, wild-type enzyme; wild-type, pH 6.5, 37°C
0.3
NADH
mutant K271R/N273, pH 6.5, 37°C; pH 6.5, 37°C, mutant enzyme K271R/N273D
0.244
NADPH
pH 6.5, 37°C, wild-type enzyme; wild-type, pH 6.5, 37°C
0.747
NADPH
mutant K271R/N273, pH 6.5, 37°C; pH 6.5, 37°C, mutant enzyme K271R/N273D
3.9
pentanal
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
14.7
pentanal
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
13.2
propionaldehyde
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
78
propionaldehyde
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
additional information
additional information
-
KM-value determined with cell extract
-
additional information
additional information
-
KM-values determined with crude extracts of native enzyme, mutant enzyme K270M and mutant enzyme K270R
-
additional information
additional information
Michaelis-Menten kinetics
-
additional information
additional information
-
kinetics of wild-type and mutant enzymes, detailed overview
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
6.8
2-deoxy-D-glucose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
1.4
2-deoxy-D-ribose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
5.6
L-idose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
3.5
2-deoxy-D-galactose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
8.4
2-deoxy-D-galactose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
5.4
Butanal
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
21.2
Butanal
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
24.3
D-erythrose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
27.5
D-erythrose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
10.2
D-fucose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
20.7
D-fucose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
9.4
D-galactose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
15.2
D-galactose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
8.2
D-glucose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
4.9
D-ribose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
12.2
D-ribose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
2.6
D-xylose
Scheffersomyces stipitis
-
pH 6.0, cofactor: NADPH, mutant enzyme K270S/N272P/S271G/R276F
12
D-xylose
Scheffersomyces stipitis
-
pH 6.0, cofactor: NADH, mutant enzyme K270S/N272P/S271G/R276F
16.9
D-xylose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
23.5
D-xylose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
25110
D-xylose
Rasamsonia emersonii
C5J3R6
pH 6.5, 37°C, cosubstrate: NADH, mutant enzyme K271R/N273D
100900
D-xylose
Rasamsonia emersonii
C5J3R6
pH 6.5, 37°C, cosubstrate: NADPH, mutant enzyme K271R/N273D
324000
D-xylose
Rasamsonia emersonii
C5J3R6
pH 6.5, 37°C, cosubstrate: NADPH, wild-type enzyme
14.1
DL-glyceraldehyde
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
24.3
DL-glyceraldehyde
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
13.5
L-arabinose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
24.5
L-arabinose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
6.6
L-Lyxose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
18.4
L-Lyxose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
5.9
pentanal
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
20.7
pentanal
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
4.6
propionaldehyde
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
6.9
propionaldehyde
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
117
2-deoxy-D-glucose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
531
36.8
2-deoxy-D-ribose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
2811
3.7
2-deoxy-D-galactose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
2006
6.3
2-deoxy-D-galactose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
2006
9.3
2-deoxy-D-galactose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
2006
17
2-deoxy-D-galactose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
2006
28
2-deoxy-D-galactose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
2006
38
2-deoxy-D-galactose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
2006
0.16
Butanal
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
540
0.93
Butanal
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
540
736
D-erythrose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
1442
1380
D-erythrose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
1442
1457
D-fucose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
1256
2070
D-fucose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
1256
5.5
D-galactose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
71
9
D-galactose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
71
70
D-galactose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
71
249
D-galactose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
71
265
D-galactose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
71
627
D-galactose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
71
8.3
D-glucose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose; pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
35
170
D-glucose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
35
188
D-glucose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
35
833
D-glucose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
35
1370
D-glucose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
35
72
D-ribose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
292
134
D-ribose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
292
6.2
D-xylose
Scheffersomyces stipitis
-
pH 6.0, cofactor: NADPH, mutant enzyme K270S/N272P/S271G/R276F
115
6.5
D-xylose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme W23Y, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
115
9
D-xylose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
115
14
D-xylose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
115
19
D-xylose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
115
25
D-xylose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme W23F, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
115
81.7
D-xylose
Scheffersomyces stipitis
-
pH 6.0, cofactor: NADH, mutant enzyme K270S/N272P/S271G/R276F
115
680
D-xylose
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
115
1470
D-xylose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
115
1690
D-xylose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
115
0.3
DL-glyceraldehyde
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose; pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
487
5.8
DL-glyceraldehyde
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
487
8.9
DL-glyceraldehyde
Candida tenuis
-
pH 7.0, 25°C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
487
21.3
DL-glyceraldehyde
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
487
482
L-arabinose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
206
1230
L-arabinose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
206
46
L-Lyxose
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
1647
157
L-Lyxose
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
1647
59920
NADH
Rasamsonia emersonii
C5J3R6
pH 6.5, 37°C, wild-type enzyme; wild-type, pH 6.5, 37°C
8
83750
NADH
Rasamsonia emersonii
C5J3R6
mutant K271R/N273, pH 6.5, 37°C; pH 6.5, 37°C, mutant enzyme K271R/N273D
8
135500
NADPH
Rasamsonia emersonii
C5J3R6
mutant K271R/N273, pH 6.5, 37°C; pH 6.5, 37°C, mutant enzyme K271R/N273D
5
1466000
NADPH
Rasamsonia emersonii
C5J3R6
pH 6.5, 37°C, wild-type enzyme; wild-type, pH 6.5, 37°C
5
0.4
pentanal
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
781
5.31
pentanal
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
781
0.035
propionaldehyde
Candida intermedia
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
273
0.09
propionaldehyde
Candida intermedia
-
pH 7.0, 25°C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
273
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.16 - 0.4
-
recombinant enzyme in transgenic strains of Saccharomyces cerevisiae
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.5
6.5
7.2
additional information
-
use of response surface analysis for the maximization of xylose reductase activity as a function of pH and temperature. This methodology also makes it possible to determine a desirable working region where a high xylose reductase to xylitol dehydrogenase ratio can be attained
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4 - 6.5
pH 4.0: about 55% of maximal activity, pH 6.5: about 60% of maximal activity
4 - 7
-
pH 4.0: about 40% of maximal activity, pH 7.0: about 50% of maximal activity
4.5 - 7.5
-
pH 4.5: about 55% of maximal activity, pH 7.5: about 50% of maximal activity
5 - 7
pH 5.0: 81% of maximal activity, pH 7.0: about 65% of maximal activity
5 - 8
5 - 8
; active from pH 5 to pH 8, wild-type and double-mutant (K271R/N273D) protein
5 - 8
-
the ratio of activities with NADH and NADPH is approximately constant between pH 5 and 8
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
30
37
additional information
-
use of response surface analysis for the maximization of xylose reductase activity as a function of pH and temperature. This methodology also makes it possible to determine a desirable working region where a high xylose reductase to xylitol dehydrogenase ratio can be attained
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25 - 65
-
25°C: about 55% of maximal activity, 65°C: about 65% of maximal activity
30 - 60
30 - 60
30°C: about 70% of maximal activity, 60°C: about 45% of maximal activity
30 - 60
active from 30°C to 60°C, wild-type and double-mutant (K271R/N273D) protein
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
-
the organism grows on rice straw hemicellulosic hydrolysate, as the only source of nutrient, optimization of culture conditions for production of xylitol from D-xylose, xylitol dehydrogenase remains constant, whereas the level of xylose reductase decreases when the initial xylose concentration is increased from 30 to 70 g/l, development of enzyme activities, overview
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PDB
SCOP
CATH
ORGANISM
UNIPROT
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
36000
37000
40000
x * 36000, calculated, x * 40000, SDS-PAGE; x * 40000, wild-type and double-mutant (K271R/N273D) histidine-tagged protein, SDS-PAGE
58000
-
D-xylose reductase 1, D-xylose reductase 2, D-xylose reductase 3, gel filtration
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
?
x * 36000, calculated, x * 40000, SDS-PAGE; x * 40000, wild-type and double-mutant (K271R/N273D) histidine-tagged protein, SDS-PAGE
dimer
-
2 * 364978, D-xylose reductase 1, ion-spray mass spectrometry; 2 * 365540, D-xylose reductase 2, ion-spray mass spectrometry
dimer
-
2 * 364978, D-xylose reductase 1, ion-spray mass spectrometry; 2 * 365540, D-xylose reductase 2, ion-spray mass spectrometry
-
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
the purified N309D mutant is crystallized by the hanging-drop vapour-diffusion method at 25°C. The best-diffracting crystals are grown using a well solution consisting of 2.1 M (NH4)2SO4, 100 mM sodium acetate and 100 mM sodium citrate, pH 6.4. 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
-
hanging-drop vapour diffusion method, to 2.91 A resolution. Unit cell belongs to space group P31 or P32, presence of four XR molecules in the asymmetric unit, with 68.0% solvent content; hanging-drop vapour-diffusion method. X-ray diffraction data from xylose reductase crystals at 2.91 A resolution, the unit cell belongs to space group P3(1) or P3(2). Preliminary analysis indicates the presence of four xylose reductase molecules in the asymmetric unit, with 68.0% solvent content
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
60
additional information
-
non-ionic detergents and bovine serum albumin stabilize the enzyme to a significant extent during long-term incubation at 25°C, 30°C or 38°C
60
-
1 h, D-xylose reductase 1, D-xylose reductase 2, complete loss of activity
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
stable enzyme at 25°C in phosphate and Tris buffer of various ionic strengths between pH 6.0 and 7.0
-
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OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
identification of the most suitable operating conditions for purification, the aqueous two-phase systems proves effective for partial purification of xylose reductase in cell-free crude extract
-
recombinant enzyme
recombinant His-tagged enzyme from Escherichia coli strain UT5600 by nickel affinity chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
construction of Kluyveromyces marxianus strains expressing Pichia stipitis Psxyl1 genes leading to reversed cofactor specificity and ethanol and xylitol production
-
expressed in Escherichia coli BL21 (DE3), subloned into the pYES2 vector and transformed into Saccharomyces cerevisiae W303-1A
expression in Candida tropicalis increases production of ethanol and glycerol
expression in Escherichia coli
expression in Escherichia coli as a His6-tagged fusion protein
expression in Escherichia coli as a His6-tagged fusion protein in high yield
-
expression in Escherichia coli; expression of histidine-tagged wild type Texr and mutant Texr K271R/N273D in Escherichia coli BL21-Star DE3
expression in Escherichia coli; subcloned into the expression plasmid pET28a(+) and subsequently transformed into Escherichia coli BL21(DE3)pLysS cells for recombinant protein expression
expression of single-site mutant xylose reductase from Candida tenuis (CtXR (K274R)) results in recombinant Corynebacterium glutamicum strain CtXR4 that produces 26.5 g/l xylitol at 3.1 g/l*h. To eliminate possible formation of toxic intracellular xylitol phosphate, genes encoding xylulokinase (XylB) and phosphoenolpyruvate-dependent fructose phosphotransferase (PTSfru) are disrupted to yield strain CtXR7
-
expression of the mutant xylulose reductase from Candida tenuis in Saccharomyces cerevisiae, co-expression with an engineered xylitol dehydrogenase, with altered cofactor specificity, from Galactocandida mastotermitis, the transformed strain shows up to 50% decreased glycerol yield without increase in ethanol during xylose fermentation, overview
-
gene XYL1
gene ZMO0976, expression of His-tagged enzyme in Escherichia coli strain UT5600
overexpression of wild-type enzyme and mutant enzyme, with modified cofactor specificity, in Saccharomyces cerevisiae strain CEN.PK 113-7D under control of the constitutive TDH3 promoter, coexpression with xylitol dehydrogenase from Galactocandida mastotermitis, The strain harboring the xylose reductase double mutant shows 42% enhanced ethanol yield compared to the reference strain harboring wild-type xylose reductase during anaerobic bioreactor conversions of xylose. Likewise, the yields of xylitol and glycerol are decreased by 52% and 57% respectively in the xylose reductase mutant strain
-
the cofactor preference of Pichia stipitis xylose reductase is altered by site-directed mutagenesis. When the K270R xylose reductase is combined with a metabolic engineering strategy that ensures high xylose utilization capabilities, a recombinant Saccharomyces cerevisiae strain is created that provides a unique combination of high xylose consumption rate, high ethanol yield and low xylitol yield during ethanolic xylose fermentation
-
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
formation of xylose reductase in the yeast Candida tropicalis is significantly repressed in cells grown on medium that contains glucose as carbon and energy source, because of the repressive effect of glucose
the Aspergillus niger transcriptional activator XlnR, which is involved in the degradation of the polysaccharides xylan and cellulose, also regulates D-xylose reductase gene expression
formation of xylose reductase in the yeast Candida tropicalis is significantly repressed in cells grown on medium that contains glucose as carbon and energy source, because of the repressive effect of glucose
-
formation of xylose reductase in the yeast Candida tropicalis is significantly repressed in cells grown on medium that contains glucose as carbon and energy source, because of the repressive effect of glucose
-
-
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
H113A
-
mutation causes a 10000-100000fold decrease in the rate constant for hydride transfer from NADH to 9,10-phenanthrenequinone, whose value in the wild-type enzyme is about 800 per s
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
L80A
-
mutation causes a 10000-100000fold decrease in the rate constant for hydride transfer from NADH to 9,10-phenanthrenequinone, whose value in the wild-type enzyme is about 800 per s
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 8–9 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 8–9 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
Y51A
-
mutation causes a 10000-100000fold decrease in the rate constant for hydride transfer from NADH to 9,10-phenanthrenequinone, whose value in the wild-type enzyme is about 800 per s
K271R/N273D
catalytic efficiency of the double mutant increases 1.4fold with NADH and decreases 10.8fold with NADPH relative to the wild-type enzyme; in the double mutant, affinity for NADPH decreases 3.1fold, while affinity for NADH remains relatively unchanged in comparison with the wild-type enzyme. The turnover number increases 1.6fold for the double mutant with NADH and decreases 3.2fold with NADPH relative to the wild-type enzyme. As a consequence, the catalytic efficiency of the double mutant (kcat/Km) increases 1.4fold with NADH and decreases 10.8fold with NADPH relative to the wild-type enzyme. Using the specificity constant (kcat/Km (NADH)/kcat/Km(NADPH)) the coenzyme preference for NADH is improved 16fold in the TeXR K271R/N273D double-mutant enzyme
K274M
-
site-directed mutagenesis, the mutant enzyme shows increased activity and altered kinetics compared to the wild-type enzyme
K74M/N276D
-
site-directed mutagenesis, the mutant enzyme shows increased activity and altered kinetics compared to the wild-type enzyme
N272D
-
site-directed mutagenesis, results in strain TMB 3422, the mutation enables the yeast for anaerobic growth on xylose displayed higher aerobic growth rates
N272D/P275Q
-
site-directed mutagenesis, results in strain TMB 3421, the mutations enable the yeast for anaerobic growth on xylose displayed higher aerobic growth rates
N276D
-
site-directed mutagenesis, the mutant enzyme shows increased activity and altered kinetics compared to the wild-type enzyme
P275Q
-
site-directed mutagenesis, results in strain TMB 3423, the mutation enables the yeast for anaerobic growth on xylose displayed higher aerobic growth rates
N272D
-
site-directed mutagenesis, results in strain TMB 3422, the mutation enables the yeast for anaerobic growth on xylose displayed higher aerobic growth rates
-
N272D/P275Q
-
site-directed mutagenesis, results in strain TMB 3421, the mutations enable the yeast for anaerobic growth on xylose displayed higher aerobic growth rates
-
P275Q
-
site-directed mutagenesis, results in strain TMB 3423, the mutation enables the yeast for anaerobic growth on xylose displayed higher aerobic growth rates
-
K21A/N272D
catalytic efficiency is almost 9fold that of the K21A mutant and 2fold that of the wild-type enzyme. Strong preference for NADH over NADPH
K270M
-
mutation results in a significant increase in the Km values for both NADPH and NADH. The kinetic parameters for the NADH-linked reaction catalyzed by the K270M mutant could not even be determined since this mutant could not be saturated with NADH
K270R
-
mutation increases the Km value for NADPH 25fold, while the Km for NADH only increased two-fold
K270S/N272P/S271G/R276F
-
the mutant shows a 25fold preference toward NADH over NADPH by a factor of about 13fold, or an improvement of about 42fold, as measured by the ratio of the specificity constant kcat/Km coenzyme. Compared with the wild-type, the kcat(NADH) is slightly lower, while the kcat(NADPH) decreases by a factor of about 10
additional information
additional information
-
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
-
xylitol production is increased by expression of codon-optimized Neurospora crassa xylose reductase gene under control of a constitutive glyceraldehyde-3-phosphate dehydrogenase promoter in a Candida tropicalis xylitol dehydrogenase gene (XYL2)-disrupted strain resulting in recombinant strain LNG2, overview
additional information
-
xylitol production is increased by expression of codon-optimized Neurospora crassa xylose reductase gene under control of a constitutive glyceraldehyde-3-phosphate dehydrogenase promoter in a Candida tropicalis xylitol dehydrogenase gene (XYL2)-disrupted strain resulting in recombinant strain LNG2, overview
-
additional information
-
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
-
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
-
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
-
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
-
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
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Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
denaturation buffers of either pH 6.0 or 8.0, containing urea in concentrations of 2, 4, 6, and 8 M, are used and analysed in SDS-PAGE. Optimal solvation of the XylR giving the lowest background of Escherichia coli proteins is performed with 4 M urea at pH 8.0. For renaturation, a set of buffers containing 0, 0.1, 0.5, 1 and 1.5 mM glutathione (red:ox = 1:1) at pH values of 5.0, 6.0, 7.0, and 8.0 are tested. Refolding occurrs at 8°C and its progress is analysed by assaying the volumetric activity in the respective buffers. Best renaturation results are obtained in a 20 mM Tris/HCl buffer at pH 7.0 without glutathione. After 4 days about 70% of the activity of the XylR is recovered. Buffers at pH 8.0 work slightly less efficient compared to that of pH 7.0. At pH 5.0 and 6.0 refolding is drastically reduced. Increasing concentrations of glutathione do not improve renaturation
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
synthesis
synthesis
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this enzyme is one of the most active xylose reductases and may be used for the in vitro production of xylitol
synthesis
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the cofactor preference of Pichia stipitis xylose reductase is altered by site-directed mutagenesis. When the K270R xylose reductase is combined with a metabolic engineering strategy that ensures high xylose utilization capabilities, a recombinant Saccharomyces cerevisiae strain is created that provides a unique combination of high xylose consumption rate, high ethanol yield and low xylitol yield during ethanolic xylose fermentation
synthesis
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dual specific xylose reductase (dsXR) has an about 4fold higher specificity for NADH than NADPH. This fact could make this enzyme an interesting candidate to be used in metabolic engineering of the yeast xylose metabolism, likely in Saccharomyces cerevisiae. Increased levels of dsXR activity could contribute to an improvement of ethanol production from D-xylose by reducing the cofactor imbalance of the initial catabolic pathway
synthesis
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the enzyme is useful for xylitol bioproduction, profiles, overview
synthesis
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D-xylose is the second most abundant renewable sugar in nature, and its fermentation to ethanol has great economical potential. Unfortunately, Saccharomyces cerevisiae, which has been optimized for ethanol production, cannot utilize xylose efficiently, while D-xylulose, an isomerization product of D-xylose, can be assimilated. A major strategy for constructing xylose-fermenting Saccharomyces cerevisiae is to introduce genes involved in xylose metabolism from other organisms. Xylose reductase and xylitol dehydrogenase (EC 1.1.1.9) from the xylose-fermenting yeast Pichia stipitis are cloned into Saccharomyces cerevisiae to allow xylose fermentation to ethanol. In this case, xylose is converted into xylulose by the sequential actions of two oxidoreductases. First, Pichia stipitis xylose reductase catalyses the reduction of xylose into xylitol with NAD(P)H as co-substrate. Xylitol is then oxidized by PsXDH (Pichia stipitis xylitol dehydrogenase) which uses NAD+ exclusively as co-substrate to yield xylulose. The different coenzyme specificity of the two enzymes xylose reductase and xylitol dehydrogenase, however, creates an intracellular redox imbalance, which results in low ethanol yields and considerable xylitol by-product formation. A mutant is constructed that shows an altered active site that is more unfavorable for NADPH than NADH in terms of both Km and kcat. There are potentials for application of the mutant (K270S/N272P/S271G/R276F) in constructing a more balanced xylose reductase/xylitol dehydrogenase pathway in recombinant xylose-fermenting Saccharomyces cerevisiae strains
synthesis
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fermentation of mixed glucose-xylose substrates in Saccharomyces cerevisiae strains BP10001 and BP000, expressing Candida tenuis xylose reductase in mutated NADH-preferring form and NADPH-preferring wild-type form, respectively. Glucose and xylose, each at 10 g/l, are converted sequentially. The distribution of fermentation products from glucose is identical for both strains whereas when using xylose, BP10001 shows enhanced ethanol yield and decreased yields of xylitol and glycerol as compared to BP000. Increase in xylose concentration from 10 to 50 g/l results in acceleration of substrate uptake by BP10001 and reduction of the xylitol yield. In mixed substrate batches, xylose is taken up at low glucose concentrations and up to 5fold enhanced xylose uptake rate is found towards glucose depletion
synthesis
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the enzyme is useful for xylitol bioproduction, profiles, overview
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synthesis
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the enzyme is useful for xylitol bioproduction, profiles, overview
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LINKS TO OTHER DATABASES (specific for EC-Number 1.1.1.307)
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