Information on EC 1.1.1.307 - D-xylose reductase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
1.1.1.307
-
RECOMMENDED NAME
GeneOntology No.
D-xylose reductase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
xylitol + NAD(P)+ = D-xylose + NAD(P)H + H+
show the reaction diagram
-
-
-
-
xylitol + NAD(P)+ = D-xylose + NAD(P)H + H+
show the reaction diagram
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
-
xylitol + NAD(P)+ = D-xylose + NAD(P)H + H+
show the reaction diagram
ordered bi-substrate mechanism in which the coenzyme binds first
-
xylitol + NAD+ = D-xylose + NADH + H+
show the reaction diagram
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
-
xylitol + NAD+ = D-xylose + NADH + H+
show the reaction diagram
-
-
-
-
xylitol + NADP+ = D-xylose + NADPH + H+
show the reaction diagram
-
-
-
-
xylitol + NADP+ = D-xylose + NADPH + H+
show the reaction diagram
kinetic mechanism of xylose reductase is iso-ordered bi bi
-
PATHWAY
KEGG Link
MetaCyc Link
Metabolic pathways
-
Pentose and glucuronate interconversions
-
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.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
CtXR
O74237
-
CtXR
C1K8Y9
-
D-xylose reductase
-
-
D-xylose reductase 1
-
-
D-xylose reductase 1
Candida tropicalis IF0 0618
-
-
-
D-xylose reductase 2
-
-
D-xylose reductase 2
Candida tropicalis IF0 0618
-
-
-
D-xylose reductase 3
-
-
D-xylose reductase 3
Candida tropicalis IF0 0618
-
-
-
dsXR
-
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)
msXR
-
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)
NAD(P)H-dependent xylose reductase
-
-
NAD(P)H-dependent xylose reductase
-
-
NADPH-preferring xylose reductase
-
-
NADPH-preferring xylose reductase
Kluyveromyces marxianus YHJ010
-
-
-
XR1
Candida tropicalis IF0 0618
-
-
-
XR2
Candida tropicalis IF0 0618
-
-
-
XYL1
Candida parapsilosis KFCC-10875
Q6Y0Z3
-
-
XYL1
C1K8Y9
gene name
xylose reductase
-
-
xylose reductase
-
-
xylose reductase
-
-
xylose reductase
Candida tropicalis ATCC 20336
-
-
-
xylose reductase
-
-
xylose reductase
Kluyveromyces marxianus YHJ010
-
-
-
xylose reductase
Meyerozyma guilliermondii FTI, Meyerozyma guilliermondii FTI 20037
-
-
-
xylose reductase
-
-
xylose reductase
F2YCN5
-
xylose reductase
Zymomonas mobilis ZM4
F2YCN5
-
-
XylR
O74237
-
XylR
Candida tenuis CBS 4435
O74237
-
-
XyrA
Q9P8R5
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
strain F-3
-
-
Manually annotated by BRENDA team
Candida diddensiae F-3
strain F-3
-
-
Manually annotated by BRENDA team
KFCC-10875
SwissProt
Manually annotated by BRENDA team
Candida parapsilosis KFCC-10875
KFCC-10875
SwissProt
Manually annotated by BRENDA team
strain Y-1632
-
-
Manually annotated by BRENDA team
Candida shehatae Y-1632
strain Y-1632
-
-
Manually annotated by BRENDA team
strain VGI-II
-
-
Manually annotated by BRENDA team
Candida silvanorum VGI-II
strain VGI-II
-
-
Manually annotated by BRENDA team
strain CBS 4435
SwissProt
Manually annotated by BRENDA team
Candida tenuis CBS 4435
strain CBS 4435
SwissProt
Manually annotated by BRENDA team
IF0 0618
-
-
Manually annotated by BRENDA team
SCTCC 300249
UniProt
Manually annotated by BRENDA team
strain Y-456
-
-
Manually annotated by BRENDA team
Candida tropicalis ATCC 20336
-
-
-
Manually annotated by BRENDA team
Candida tropicalis IF0 0618
IF0 0618
-
-
Manually annotated by BRENDA team
Candida tropicalis SCTCC 300249
SCTCC 300249
UniProt
Manually annotated by BRENDA team
Candida tropicalis Y-456
strain Y-456
-
-
Manually annotated by BRENDA team
Corynebacterium glutamicum ATCC13032
gene XYL1
-
-
Manually annotated by BRENDA team
strain UFV-170 XR
-
-
Manually annotated by BRENDA team
Debaryomyces hansenii UFV-170 XR
strain UFV-170 XR
-
-
Manually annotated by BRENDA team
Kluyveromyces marxianus Y-488
Y-488
-
-
Manually annotated by BRENDA team
Kluyveromyces marxianus YHJ010
-
-
-
Manually annotated by BRENDA team
FTI 20037, ATCC 201935
-
-
Manually annotated by BRENDA team
strain Y-1017
-
-
Manually annotated by BRENDA team
Meyerozyma guilliermondii FTI 20037
FTI 20037
-
-
Manually annotated by BRENDA team
Meyerozyma guilliermondii Y-1017
strain Y-1017
-
-
Manually annotated by BRENDA team
strain Y-1532, strain Y-1533, strain Y-1634
-
-
Manually annotated by BRENDA team
Saccharomyces cerevisiae TMB 3420
-
-
-
Manually annotated by BRENDA team
strain Y-2160
-
-
Manually annotated by BRENDA team
Scheffersomyces stipitis Y-2160
strain Y-2160
-
-
Manually annotated by BRENDA team
Torulopsis molishiama
strain 55
-
-
Manually annotated by BRENDA team
Torulopsis molishiama 55
strain 55
-
-
Manually annotated by BRENDA team
ssp. mobilis, and strain A3, an engineered strain ZM4 that is adapted to 5% D-xylose, gene ZMO0976
UniProt
Manually annotated by BRENDA team
Zymomonas mobilis ZM4
ssp. mobilis, and strain A3, an engineered strain ZM4 that is adapted to 5% D-xylose, gene ZMO0976
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
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
-
key enzymes for xylitol production in yeasts are xylose reductase and xylitol dehydrogenase, EC 1.1.1.9, overview
metabolism
-
xylose reductase is the first enzyme in D-xylose metabolism, catalyzing the reduction of D-xylose to xylitol
metabolism
Candida diddensiae F-3, Candida shehatae Y-1632, Candida silvanorum VGI-II
-
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
Candida tropicalis ATCC 20336
-
xylose reductase is the first enzyme in D-xylose metabolism, catalyzing the reduction of D-xylose to xylitol
-
metabolism
Candida tropicalis Y-456, Kluyveromyces marxianus Y-488, Meyerozyma guilliermondii Y-1017, Scheffersomyces stipitis Y-2160, 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
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-deoxy-D-galactose + NADH
?
show the reaction diagram
O74237
-
-
-
?
2-deoxy-D-galactose + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
2-deoxy-D-galactose + NADPH + H+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
2-deoxy-D-glucose + NADPH + H+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
2-deoxy-D-ribose + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
2-deoxy-D-ribose + NADPH + H+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
5-hydroxymethylfurfural + NADH + H+
5-hydroxyfurfurol + NAD+
show the reaction diagram
Zymomonas mobilis, Zymomonas mobilis ZM4
F2YCN5
low activity
-
-
r
9,10-phenanthrenequinone + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
-
?
acetaldehyde + NADH + H+
ethanol + NAD+
show the reaction diagram
Zymomonas mobilis, Zymomonas mobilis ZM4
F2YCN5
-
-
-
r
benzaldehyde + NADH + H+
benzyl alcohol + NAD+
show the reaction diagram
Zymomonas mobilis, Zymomonas mobilis ZM4
F2YCN5
best substrate
-
-
r
butanal + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
butanal + NADPH + H+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-arabinose + NADPH + H+
?
show the reaction diagram
Candida tropicalis, Candida tropicalis SCTCC 300249
A9QVV8
about 60% of the activity compared to D-xylose
-
-
?
D-arabinose + NADPH + H+
D-arabinitol + NADP+
show the reaction diagram
Candida parapsilosis, Candida parapsilosis KFCC-10875
Q6Y0Z3
-
-
-
?
D-erythrose + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
D-erythrose + NADPH + H+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-erythrose + NADPH + H+
D-erythritol + NADP+
show the reaction diagram
Candida parapsilosis, Candida parapsilosis KFCC-10875
Q6Y0Z3
-
-
-
?
D-erythrose + NADPH + H+
erythritol + NADP+
show the reaction diagram
-
catalytic efficiency is 100fold higher than the catalytic efficiency for D-xylose
-
-
?
D-fucose + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
D-fucose + NADPH + H+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-galactose + NADH
?
show the reaction diagram
O74237
-
-
-
?
D-galactose + NADH + H+
?
show the reaction diagram
-
-
-
-
?
D-galactose + NADH + H+
?
show the reaction diagram
A0MTG4
48% of the activity compared to D-xylose (with NADH as cofactor)
-
-
?
D-galactose + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
D-galactose + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
D-galactose + NADPH + H+
?
show the reaction diagram
A9QVV8
about 70% of the activity compared to D-xylose
-
-
?
D-galactose + NADPH + H+
?
show the reaction diagram
-
catalytic efficiency is 9.1% of the catalytic efficiency for D-xylose
-
-
?
D-galactose + NADPH + H+
?
show the reaction diagram
-
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+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-galactose + NADPH + H+
?
show the reaction diagram
Candida tropicalis IF0 0618
-
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-glucose + NADH
?
show the reaction diagram
O74237
-
-
-
?
D-glucose + NADH + H+
?
show the reaction diagram
-
-
-
-
?
D-glucose + NADH + H+
?
show the reaction diagram
A0MTG4
10 of the activity compared to D-xylose (with NADH as cofactor)
-
-
?
D-glucose + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
D-glucose + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
D-glucose + NADPH + H+
?
show the reaction diagram
A9QVV8
about 15% of the activity compared to D-xylose
-
-
?
D-glucose + NADPH + H+
?
show the reaction diagram
-
catalytic efficiency is 3.3% of the catalytic efficiency for D-xylose
-
-
?
D-glucose + NADPH + H+
?
show the reaction diagram
-
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+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-glucose + NADPH + H+
?
show the reaction diagram
Candida tropicalis IF0 0618
-
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-glucosone + NADPH + H+
D-fructose + NADP+
show the reaction diagram
-
catalytic efficiency is 22fold higher than the catalytic efficiency for D-xylose
-
-
?
D-glyceraldehyde + NADH
?
show the reaction diagram
O74237
-
-
-
?
D-lyxose + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
D-lyxose + NADPH + H+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-mannose + NADH + H+
?
show the reaction diagram
A0MTG4
8% of the activity compared to D-xylose (with NADH as cofactor)
-
-
?
D-ribose + NADH + H+
?
show the reaction diagram
-
-
-
-
?
D-ribose + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
D-ribose + NADPH + H+
?
show the reaction diagram
-
-
-
-
?
D-ribose + NADPH + H+
?
show the reaction diagram
Q6Y0Z3
-
-
-
?
D-ribose + NADPH + H+
?
show the reaction diagram
A9QVV8
about 90% of the activity compared to D-xylose
-
-
?
D-ribose + NADPH + H+
?
show the reaction diagram
-
catalytic efficiency is 41% of the catalytic efficiency for D-xylose
-
-
?
D-ribose + NADPH + H+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
D-ribose + NADPH + H+
?
show the reaction diagram
Candida parapsilosis KFCC-10875
Q6Y0Z3
-
-
-
?
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
show the reaction diagram
-
-
-
-
?
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
show the reaction diagram
-
-
-
-
?
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
show the reaction diagram
-
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)+
show the reaction diagram
Meyerozyma guilliermondii FTI
-
-, 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)+
show the reaction diagram
Meyerozyma guilliermondii FTI 20037
-
-, 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+
show the reaction diagram
-
-
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
-
-
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
-
-
-
-
r
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
O74237
-
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
C5J3R6
-
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
A9QVV8
-
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
P31867
-
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
F2YCN5
-
-
-
r
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
A0MTG4
NADPH is the preferred cofactor
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
-
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+
show the reaction diagram
-
aldehyde reduction is favoured
-
-
r
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
-
catalytic efficiency for NADPH is more than 100fold higher than the catalytic efficiency for NADH
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
-
dual (NADH and NADPH) coenzyme specificity
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
A9QVV8
dual coenzyme specificity, Km for NADPH: 0.0455 mM, Km for NADH: 0.162 mM
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
O74237
kcat of wilde-type enzyme increases by a factor of 1.73 when NADPH replaces NADH
-
-
r
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
-
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+
show the reaction diagram
Q6Y0Z3
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+
show the reaction diagram
-
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+
show the reaction diagram
-
wild-type enzyme prefers NADPH over NADH
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
C5J3R6
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+
show the reaction diagram
Zymomonas mobilis ZM4
F2YCN5
-
-
-
r
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
Candida parapsilosis KFCC-10875
Q6Y0Z3
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+
show the reaction diagram
Candida tropicalis SCTCC 300249
A9QVV8
-, dual coenzyme specificity, Km for NADPH: 0.0455 mM, Km for NADH: 0.162 mM
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
Candida tenuis CBS 4435
O74237
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
O74237
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
C1K8Y9
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
C5J3R6
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
Q6Y0Z3
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
P31867
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
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+
show the reaction diagram
A9QVV8
key enzyme in xylose metabolism
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
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+
show the reaction diagram
-
xylose reductase is one of the key enzymes for xylose fermentation
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
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+
show the reaction diagram
-
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+
show the reaction diagram
-
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+
show the reaction diagram
-
catalytic efficiency for NADPH is more than 100fold higher than the catalytic efficiency for NADH
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
dual (NADH and NADPH) coenzyme specificity
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
A9QVV8
dual coenzyme specificity, Km for NADPH: 0.0455 mM, Km for NADH: 0.162 mM
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
O74237
kcat of wild-type enzyme increases by a factor of 1.73 when NADPH replaces NADH
-
-
r
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
kinetic mechanism of xylose reductase is iso-ordered bi bi
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
A0MTG4
NADPH is the preferred cofactor, specific for D-xylose
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
reaction is catalyzed by NADPH-dependent monospecific xylose reductase (msXR), and by dual specific xylose reductase (dsXR)
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
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+
show the reaction diagram
-
wild-type enzyme prefers NADPH over NADH
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
C5J3R6
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+
show the reaction diagram
Meyerozyma guilliermondii, Candida tropicalis, Kluyveromyces marxianus, Pachysolen tannophilus, Candida parapsilosis, Scheffersomyces stipitis, Candida shehatae, Candida intermedia, Candida diddensiae, Candida silvanorum, Torulopsis molishiama, Kluyveromyces marxianus Y-488, Candida silvanorum VGI-II, Meyerozyma guilliermondii Y-1017, Candida diddensiae F-3
-
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+
show the reaction diagram
Candida parapsilosis KFCC-10875
Q6Y0Z3
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
Candida tropicalis IF0 0618
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
Candida shehatae Y-1632, Candida tropicalis Y-456
-
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+
show the reaction diagram
Candida tropicalis SCTCC 300249
A9QVV8
key enzyme in xylose metabolism, dual coenzyme specificity, Km for NADPH: 0.0455 mM, Km for NADH: 0.162 mM
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
Torulopsis molishiama 55, Scheffersomyces stipitis Y-2160
-
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+
show the reaction diagram
Corynebacterium glutamicum ATCC13032
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
Candida tenuis CBS 4435
O74237
-, xylose reductases catalyse the initial reaction in the xylose utilisation pathway, the NAD(P)H dependent reduction of xylose to xylitol
-
-
?
DL-glyceraldehyde + NADH + H+
glycerol + NAD+
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
DL-glyceraldehyde + NADH + H+
glycerol + NAD+
show the reaction diagram
-
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+
show the reaction diagram
-
catalytic efficiency is 37fold higher than the catalytic efficiency for D-xylose
-
-
?
DL-glyceraldehyde + NADPH + H+
glycerol + NADP+
show the reaction diagram
-
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+
show the reaction diagram
-
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+
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
DL-glyceraldehyde + NADPH + H+
glycerol + NADP+
show the reaction diagram
Candida tropicalis IF0 0618
-
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
-
-
?
furfural + NADH + H+
furfurol + NAD+
show the reaction diagram
Zymomonas mobilis, Zymomonas mobilis ZM4
F2YCN5
-
-
-
r
L-arabinose + NADH + H+
arabinitol + NAD+
show the reaction diagram
-
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+
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
L-arabinose + NADPH + H+
arabinitol + NADP+
show the reaction diagram
-
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 + NADPH + H+
L-arabinitol + NADP+
show the reaction diagram
-
catalytic efficiency is 2fold higher than the catalytic efficiency for D-xylose
-
-
?
L-arabinose + NADPH + H+
L-arabinitol + NADP+
show the reaction diagram
-
catalytic efficiency is 41% of the catalytic efficiency for D-xylose
-
-
?
L-arabinose + NADPH + H+
L-arabinitol + NADP+
show the reaction diagram
-
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+
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
L-arabinose + NADPH + H+
L-arabinitol + NADP+
show the reaction diagram
Candida tropicalis IF0 0618
-
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-arabitol + NADP+
show the reaction diagram
-
-
-
-
?
L-lyxose + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
L-lyxose + NADPH + H+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
pentanal + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
pentanal + NADPH + H+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
phenylglyoxal + NADPH + H+
?
show the reaction diagram
-
catalytic efficiency is 17fold higher than the catalytic efficiency for D-xylose
-
-
?
propionaldehyde + NADH + H+
?
show the reaction diagram
-
dual specific xylose reductase (dsXR)
-
-
?
propionaldehyde + NADPH + H+
?
show the reaction diagram
-
NADPH-dependent monospecific xylose reductase
-
-
?
pyridine-2-aldehyde + NADPH + H+
?
show the reaction diagram
-
catalytic efficiency is 7fold higher than the catalytic efficiency for D-xylose
-
-
?
valeraldehyde + NADPH + H+
?
show the reaction diagram
-
catalytic efficiency is 13fold higher than the catalytic efficiency for D-xylose
-
-
?
xylitol + NAD+
D-xylose + NADH + H+
show the reaction diagram
-
-
-
-
r
xylitol + NAD+
D-xylose + NADH + H+
show the reaction diagram
Candida tropicalis, Candida tropicalis ATCC 20336
-
-
-
-
r
xylitol + NAD+
D-xylose + NADH + H+
show the reaction diagram
Saccharomyces cerevisiae TMB 3420
-
-
-
-
r
xylitol + NADP+
D-xylose + NADPH + H+
show the reaction diagram
-
-
-
-
r
xylitol + NADP+
D-xylose + NADPH + H+
show the reaction diagram
-
-
-
-
r
xylitol + NADP+
D-xylose + NADPH + H+
show the reaction diagram
-
-
NADPH is the preferred cofactor
-
r
xylitol + NADP+
D-xylose + NADPH + H+
show the reaction diagram
Candida tropicalis ATCC 20336
-
-
-
-
r
xylitol + NADP+
D-xylose + NADPH + H+
show the reaction diagram
Saccharomyces cerevisiae TMB 3420
-
-
-
-
r
xylosone + NADPH + H+
?
show the reaction diagram
-
catalytic efficiency is 20fold higher than the catalytic efficiency for D-xylose
-
-
?
xylulose + NADH + H+
? + NAD+
show the reaction diagram
F2YCN5
-
-
-
r
methylglyoxal + NADPH + H+
?
show the reaction diagram
-
catalytic efficiency is 20fold higher than the catalytic efficiency for D-xylose
-
-
?
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
?
-
O74237
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
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
show the reaction diagram
-
-
-
-
?
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
show the reaction diagram
-
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)+
show the reaction diagram
Meyerozyma guilliermondii FTI, Meyerozyma guilliermondii FTI 20037
-
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+
show the reaction diagram
A9QVV8
-
-
-
?
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
Zymomonas mobilis, Zymomonas mobilis ZM4
F2YCN5
-
-
-
r
D-xylose + NADH + H+
xylitol + NAD+
show the reaction diagram
Candida tropicalis SCTCC 300249
A9QVV8
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
O74237
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
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+
show the reaction diagram
A9QVV8
key enzyme in xylose metabolism
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
-
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+
show the reaction diagram
-
xylose reductase is one of the key enzymes for xylose fermentation
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
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+
show the reaction diagram
Candida tropicalis SCTCC 300249
A9QVV8
key enzyme in xylose metabolism
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
Corynebacterium glutamicum ATCC13032
-
-
-
-
?
D-xylose + NADPH + H+
xylitol + NADP+
show the reaction diagram
Candida tenuis CBS 4435
O74237
xylose reductases catalyse the initial reaction in the xylose utilisation pathway, the NAD(P)H dependent reduction of xylose to xylitol
-
-
?
xylitol + NAD+
D-xylose + NADH + H+
show the reaction diagram
-
-
-
-
r
xylitol + NAD+
D-xylose + NADH + H+
show the reaction diagram
Candida tropicalis, Candida tropicalis ATCC 20336
-
-
-
-
r
xylitol + NAD+
D-xylose + NADH + H+
show the reaction diagram
Saccharomyces cerevisiae TMB 3420
-
-
-
-
r
xylitol + NADP+
D-xylose + NADPH + H+
show the reaction diagram
-
-
-
-
r
xylitol + NADP+
D-xylose + NADPH + H+
show the reaction diagram
-
-
-
-
r
xylitol + NADP+
D-xylose + NADPH + H+
show the reaction diagram
-
-
NADPH is the preferred cofactor
-
r
xylitol + NADP+
D-xylose + NADPH + H+
show the reaction diagram
Candida tropicalis ATCC 20336
-
-
-
-
r
xylitol + NADP+
D-xylose + NADPH + H+
show the reaction diagram
Saccharomyces cerevisiae TMB 3420
-
-
-
-
r
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
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
Q6Y0Z3
strongly prefers NADH to NADPH
NADH
-
catalytic efficiency for NADPH is more than 100fold higher than the catalytic efficiency for NADH
NADH
-
dual (NADH and NADPH) coenzyme specificity
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
A0MTG4
NADPH is the preferred cofactor
NADH
-
dual specific xylose reductase (dsXR) has an about 4fold higher specificity for NADH than NADPH
NADH
C5J3R6
catalytic efficiency is 24.5fold higher with NADPH as coenzyme than with NADH
NADH
A9QVV8
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
NADH
-
prefers NADPH as the coenzyme by about 80fold over 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
-
only NADPH-dependent xylose reductase is obtained under the cultivation conditions
NADPH
Q6Y0Z3
strongly prefers NADH to NADPH
NADPH
-
catalytic efficiency for NADPH is more than 100fold higher than the catalytic efficiency for NADH
NADPH
-
dual (NADH and NADPH) coenzyme specificity
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
-
specific for NADPH
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
A0MTG4
NADPH is the preferred cofactor
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
C5J3R6
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
-
no activity with NADH
NADPH
A9QVV8
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
NADPH
-
prefers NADPH as the coenzyme by about 80fold over NADH
NADPH
-
preferred cofactor
NADPH
-
preferred cofactor
additional information
-
Kluyveromyces marxianus strains expressing Pichia stipitis Psxyl1 genes show reversed cofactor specificity, overview
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
CaCl2
Q6Y0Z3
1 mM, stimulates
CoCl2
Q6Y0Z3
1 mM, stimulates
FeCl2
Q6Y0Z3
1 mM, stimulates
Li+
A0MTG4
Ca2+, Li+, Mg2+, Mn2+ and NH4+ at 10 mM decrease activity by 10-50%
Mg2+
A0MTG4
Ca2+, Li+, Mg2+, Mn2+ and NH4+ at 10 mM decrease activity by 10-50%
MgCl2
Q6Y0Z3
1 mM, stimulates
Mn2+
A0MTG4
Ca2+, Li+, Mg2+, Mn2+ and NH4+ at 10 mM decrease activity by 10-50%
NH4+
A0MTG4
Ca2+, Li+, Mg2+, Mn2+ and NH4+ at 10 mM decrease activity by 10-50%
NiCl2
Q6Y0Z3
1 mM, stimulates
ZnCl2
Q6Y0Z3
1 mM, stimulates
MnCl2
Q6Y0Z3
1 mM, stimulates
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
Q6Y0Z3
neither inhibited nor activated by EDTA at concentrations ranging from 1 to 10 mM
additional information
-
no effect: 1 mM CuSO4
additional information
-
no requirement for divalent cation is observed
additional information
A9QVV8
no increase in activity in presence of 1 mM NaCl and 1 mM MgSO4
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
AMP
-
2 mM, completely abolishes D-xylose reduction
ATP
-
2 mM, completely abolishes D-xylose reduction, competitive
cholic acid
-
0.1% (w/v), 30% inhibition
Cu2+
Q6Y0Z3
activity is completely restored by addition of EDTA
deoxycholic acid
-
0.1% (w/v), 30% inhibition
dithiothreitol
-
1 mM, 40% inhibition
dithiothreitol
-
1 mM, 25% inhibition
DTT
A0MTG4
1 mM, 35% inhibition
EDTA
-
1 mM, 30% inhibition
Hg2+
-
0.001 mM, 2 min, complete inhibition
N-bromosuccinimide
-
NADPH protects
NaCl
-
50 mM, 25% inhibition
NAD+
Q6Y0Z3
competitive with NADH, non-competitive with 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+
-
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
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
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
p-chloromercuribenzoate
-
0.001 mM, 2 min, complete inhibition
pyridoxal 5'-phosphate
-
gradual inactivation. NADH, ATP or 2'-AMP protects. No protection by D-xylose
sodium phosphite
-
200 mM, 37% inhibition
xylitol
Q6Y0Z3
non-competitive against NADH and D-xylose
Zn2+
-
25 mM, 95% inhibition
Mn2+
-
25 mM, 95% inhibition
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 0.1% w/v sodium azide
-
additional information
-
no effect by 5 mM EDTA, 500 mM sulfate, 5 mM 2-mercaptoethanol
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-mercaptoethanol
Q6Y0Z3
1 mM, increases activity by 13%
Bovine serum albumin
-
0.1% (w/v), 10-15% activation
-
Bovine serum albumin
-
1 mg/ml, 30% activation
-
Bovine serum albumin
A0MTG4
1 mg/ml. the specific activity is increased by 20-30%
-
Bovine serum albumin
A9QVV8
1 mg/ml, 20% increase of activity
-
cysteine
Q6Y0Z3
1 mM, increases activity by 29%
dithiothreitol
Q6Y0Z3
1 mM, increases activity by 43%
DTT
A9QVV8
1 mM, 27% increase of activity
EDTA
-
1 mM, 5% activation
EDTA
A0MTG4
1 mM, 30% activation
Triton X-100
-
0.1% (w/v), 10-15% activation
Triton X-100
A0MTG4
0.1%, the specific activity is increased by 2030%
Tween 20
A0MTG4
0.1%, the specific activity is increased by 20-30%
Tween 80
A0MTG4
0.1%, the specific activity is increased by 20-30%
Tween-20
-
0.1% (w/v), 10-15% activation
Tween-80
-
0.1% (w/v), 10-15% activation
glutathione
Q6Y0Z3
1 mM, increases activity by 21%
additional information
Q6Y0Z3
neither inhibited nor activated by EDTA at concentrations ranging from 1 to 10 mM
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.126
-
2-deoxy-D-galactose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
0.221
-
2-deoxy-D-galactose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.058
-
2-deoxy-D-glucose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.038
-
2-deoxy-D-ribose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
1.8
-
benzaldehyde
F2YCN5
pH 7.2, temperature not specified in the publication
2
3
Butanal
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
33
-
Butanal
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
285.4
-
D-arabinose
Q6Y0Z3
pH 6.0
0.02
-
D-Erythrose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
0.033
-
D-Erythrose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
151.7
-
D-Erythrose
Q6Y0Z3
pH 6.0
0.007
-
D-fucose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
0.01
-
D-fucose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.015
-
D-galactose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
0.061
-
D-galactose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
180
-
D-galactose
-
pH 6.3, 25C
0.006
-
D-glucose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.033
-
D-glucose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
360
-
D-glucose
-
pH 6.3, 25C
0.068
-
D-ribose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
0.091
-
D-ribose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
70
-
D-ribose
-
pH 6.3, 25C
302
-
D-ribose
Q6Y0Z3
pH 6.0
0.01
-
D-xylose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
0.016
-
D-xylose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
14.8
-
D-xylose
-
pH 7.0, 22C, wild-type enzyme
24.56
-
D-xylose
C5J3R6
pH 6.5, 37C, cosubstrate: NADPH, wild-type enzyme
24.6
-
D-xylose
C5J3R6
wild-type, cosubstrate NADPH, pH 6.5, 37C
30
-
D-xylose
-
pH 7.0, D-xylose reductase 2
31.5
-
D-xylose
Q6Y0Z3
pH 6.0, coenzyme: NADH
31.5
-
D-xylose
A9QVV8
pH 5.5, 45C
33
-
D-xylose
P31867
wild-type, cosubstrate NADPH, pH 6.5, 35C
34
-
D-xylose
-
pH 6.3, 25C, cosubstrate: NADPH
34
-
D-xylose
-
pH 7.0, D-xylose reductase 3
37
-
D-xylose
-
pH 6.3, 25C, cosubstrate: NADH
37
-
D-xylose
-
pH 7.0, D-xylose reductase 1
39
-
D-xylose
-
30C
42
-
D-xylose
-
cofactor: NADH; cofactor: NADPH
67
-
D-xylose
P31867
mutant K21A/N272D, cosubstrate NADH, pH 6.5, 35C; wild-type, cosubstrate NADH, pH 6.5, 35C
72
-
D-xylose
-
pH 7, 25C, cosubstrate: NADPH
76.06
-
D-xylose
C5J3R6
pH 6.5, 37C, cosubstrate: NADH, mutant enzyme K271R/N273D
76.1
-
D-xylose
C5J3R6
mutant K271R/N273, cosubstrate NADH, pH 6.5, 37C
76.5
-
D-xylose
C5J3R6
mutant K271R/N273, cosubstrate NADPH, pH 6.5, 37C; pH 6.5, 37C, cosubstrate: NADPH, mutant enzyme K271R/N273D
78
-
D-xylose
-
pH 7.0, 25C
82
-
D-xylose
-
pH 6.0, cofactor: NADPH, wild-type enzyme
87
-
D-xylose
-
pH 7, 25C, cosubstrate: NADH
89.4
-
D-xylose
C5J3R6
pH 6.5, 37C, cosubstrate: NADH, wild-type enzyme; wild-type, cosubstrate NADH, pH 6.5, 37C
90
-
D-xylose
-
pH 6.0, cofactor: NADH, wild-type enzyme
90.44
-
D-xylose
O74237
pH 6.0, 25C, cosubstrate: NADH
96
-
D-xylose
-
pH 7.0, 25C, wild-type enzyme, with NADPH
99
-
D-xylose
-
pH 7.0, 25C, mutant N276D, with NADH
106
-
D-xylose
-
pH 7.0, 25C, mutant K274M/N276D, with NADH
142
-
D-xylose
-
pH 7.0, 25C, wild-type enzyme, with NADH
160
-
D-xylose
A0MTG4
pH 6.5, 35C
167
-
D-xylose
P31867
mutant K21A, cosubstrate NADH, pH 6.5, 35C
168
-
D-xylose
-
pH 6.0, cofactor: NADPH, mutant enzyme K270S/N272P/S271G/R276F
170
-
D-xylose
-
pH 7.0, 25C, mutant N276D, with NADPH
229
-
D-xylose
-
pH 7.0, 25C, mutant K274M, with NADH
244.3
-
D-xylose
Q6Y0Z3
pH 6.0, coenzyme: NADPH
258
-
D-xylose
F2YCN5
pH 7.2, temperature not specified in the publication
291
-
D-xylose
-
pH 6.0, cofactor: NADH, mutant enzyme K270S/N272P/S271G/R276F
506
-
D-xylose
-
pH 7.0, 25C, mutant K274M, with NADPH
722
-
D-xylose
-
pH 7.0, 25C, mutant K274M/N276D, with NADPH
1.14
-
DL-glyceraldehyde
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
2.43
-
DL-glyceraldehyde
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
4.2
-
furfural
F2YCN5
pH 7.2, temperature not specified in the publication
0.02
-
L-arabinose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.028
-
L-arabinose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
40
-
L-arabinose
-
pH 6.3, 25C
93
-
L-arabinose
-
30C
0.117
-
L-Lyxose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.027
-
NAD+
-
pH 7.0, 25C
0.0587
-
NAD+
-
pH 7, 25C
0.0033
-
NADH
Q6Y0Z3
pH 6.0
0.01
-
NADH
P31867
mutant K21A/N272D, pH 6.5, 35C
0.0106
-
NADH
-
pH 6.0, wild-type enzyme, wild-type enzyme
0.015
-
NADH
-
pH 7.0, 25C
0.016
-
NADH
-
pH 6.3, 25C
0.02
-
NADH
P31867
wild-type, pH 6.5, 35C
0.026
-
NADH
-
pH 7.0, 25C, mutant N276D
0.03
-
NADH
P31867
mutant K21A, pH 6.5, 35C
0.038
-
NADH
-
pH 7.0, 25C, wild-type enzyme
0.041
-
NADH
-
pH 7.0, 25C, mutant K274M/N276D
0.147
-
NADH
-
pH 6.0, mutant enzyme K270S/N272P/S271G/R276F
0.15
-
NADH
A0MTG4
pH 6.5, 35C
0.1619
-
NADH
A9QVV8
pH 5.5, 45C
0.254
-
NADH
-
pH 7, 25C
0.263
-
NADH
C5J3R6
pH 6.5, 37C, wild-type enzyme; wild-type, pH 6.5, 37C
0.3
-
NADH
C5J3R6
mutant K271R/N273, pH 6.5, 37C; pH 6.5, 37C, mutant enzyme K271R/N273D
0.38
-
NADH
-
pH 7.0, 25C, mutant K274M
38
-
NADH
-
pH 7.0, 25C, wild-type enzyme
40
-
NADH
-
pH 7.0, 25C, mutant enzyme D50A
0.0266
-
NADP+
-
pH 7, 25C
0.0018
-
NADPH
-
pH 6.3, 25C
0.003
-
NADPH
-
pH 7.0, 25C, wild-type enzyme
0.0048
-
NADPH
-
pH 7, 25C
0.0062
-
NADPH
-
pH 6.0, wild-type enzyme, wild-type enzyme
0.008
-
NADPH
-
30C, cosubstrate: D-xylose or L-arabinose
0.009
-
NADPH
-
pH 7.0, D-xylose reductase 3
0.014
-
NADPH
-
pH 7.0, D-xylose reductase 1
0.017
-
NADPH
-
pH 7.0, 25C, mutant N276D
0.018
-
NADPH
-
pH 7.0, D-xylose reductase 2
0.02
-
NADPH
A0MTG4
pH 6.5, 35C
0.03
-
NADPH
P31867
wild-type, pH 6.5, 35C
0.0365
-
NADPH
Q6Y0Z3
pH 6.0
0.0455
-
NADPH
A9QVV8
pH 5.5, 45C
0.075
-
NADPH
-
pH 7.0, 25C, mutant K274M
0.128
-
NADPH
-
pH 7.0, 25C, mutant K274M/N276D
0.244
-
NADPH
C5J3R6
pH 6.5, 37C, wild-type enzyme; wild-type, pH 6.5, 37C
0.427
-
NADPH
-
pH 6.0, mutant enzyme K270S/N272P/S271G/R276F
0.747
-
NADPH
C5J3R6
mutant K271R/N273, pH 6.5, 37C; pH 6.5, 37C, mutant enzyme K271R/N273D
2.3
-
NADPH
-
pH 7.0, 25C, mutant enzyme D50A
3.2
-
NADPH
-
pH 7.0, 25C, wild-type enzyme
7.6
-
NADPH
-
pH 7.0, 22C, wild-type enzyme
3.9
-
Pentanal
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
14.7
-
Pentanal
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
13.2
-
propionaldehyde
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
78
-
propionaldehyde
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
209
-
xylitol
-
pH 7.0, 25C
257
-
xylitol
-
pH 7, 25C
326.3
-
xylitol
Q6Y0Z3
pH 6.0
334
-
xylitol
-
pH 7.0, 25C, wild-type enzyme
537
-
xylitol
-
pH 7.0, 25C, mutant enzyme D50A
0.144
-
L-Lyxose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
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
F2YCN5
Michaelis-Menten kinetics
-
additional information
-
additional information
-
kinetics of wild-type and mutant enzymes, detailed overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
3.5
-
2-deoxy-D-galactose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
8.4
-
2-deoxy-D-galactose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
6.8
-
2-deoxy-D-glucose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
1.4
-
2-deoxy-D-ribose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.027
-
9,10-phenanthrenequinone
-
mutant K80A, pH 7.0, 25C
0.043
-
9,10-phenanthrenequinone
-
mutant H113A, pH 7.0, 25C
0.2
-
9,10-phenanthrenequinone
-
mutant Y51A, pH 7.0, 25C
12
-
9,10-phenanthrenequinone
-
wild-type, pH 7.0, 25C
5.4
-
Butanal
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
21.2
-
Butanal
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
24.3
-
D-Erythrose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
27.5
-
D-Erythrose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
10.2
-
D-fucose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
20.7
-
D-fucose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
9.4
-
D-galactose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
15.2
-
D-galactose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
1800
-
D-galactose
-
pH 6.3, 25C
8.2
-
D-glucose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
1320
-
D-glucose
-
pH 6.3, 25C
4.9
-
D-ribose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
12.2
-
D-ribose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
3120
-
D-ribose
-
pH 6.3, 25C
0.002
-
D-xylose
-
mutant K80A, pH 7.0, 25C
0.02
-
D-xylose
-
mutant H113A, pH 7.0, 25C
2.6
-
D-xylose
-
pH 6.0, cofactor: NADPH, mutant enzyme K270S/N272P/S271G/R276F
3
6
D-xylose
-
pH 7.0, 25C, mutant K274M, with NADPH
10
-
D-xylose
-
wild-type, pH 7.0, 25C
11
-
D-xylose
-
pH 7.0, 25C, wild-type enzyme, with NADH
12
-
D-xylose
-
pH 6.0, cofactor: NADH, mutant enzyme K270S/N272P/S271G/R276F
12
-
D-xylose
-
pH 7.0, 25C, mutant K274M/N276D, with NADH
13
-
D-xylose
-
pH 7.0, 25C, wild-type enzyme, with NADPH
13.1
-
D-xylose
-
pH 7.0, 22C, wild-type enzyme
14
-
D-xylose
-
pH 7.0, 25C, mutant N276D, with NADH
14.2
-
D-xylose
-
pH 7.0, 25C
15.4
-
D-xylose
-
pH 6.0, cofactor: NADH, wild-type enzyme
16.9
-
D-xylose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
18.11
-
D-xylose
O74237
pH 6.0, 25C, cosubstrate: NADH
18.2
-
D-xylose
-
pH 7, 25C, cosubstrate: NADH
19
-
D-xylose
-
pH 7.0, 25C, mutant K274M, with NADH
21.5
-
D-xylose
-
pH 7, 25C, cosubstrate: NADPH
23.5
-
D-xylose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
27.5
-
D-xylose
-
pH 6.0, cofactor: NADPH, wild-type enzyme
30
-
D-xylose
-
pH 7.0, 25C, mutant K274M/N276D, with NADPH
37
-
D-xylose
-
pH 7.0, 25C, mutant N276D, with NADPH
240
-
D-xylose
A9QVV8
-
310
-
D-xylose
-
pH 6.3, 25C, cosubstrate: NADH
3600
-
D-xylose
-
pH 6.3, 25C, cosubstrate: NADPH
4638
-
D-xylose
-
pH 6.3, 25C
15750
-
D-xylose
C5J3R6
pH 6.5, 37C, cosubstrate: NADH, wild-type enzyme
25110
-
D-xylose
C5J3R6
pH 6.5, 37C, cosubstrate: NADH, mutant enzyme K271R/N273D
100900
-
D-xylose
C5J3R6
pH 6.5, 37C, cosubstrate: NADPH, mutant enzyme K271R/N273D
324000
-
D-xylose
C5J3R6
pH 6.5, 37C, cosubstrate: NADPH, wild-type enzyme
14.1
-
DL-glyceraldehyde
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
24.3
-
DL-glyceraldehyde
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
13.5
-
L-arabinose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
24.5
-
L-arabinose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
1800
-
L-arabinose
-
pH 6.3, 25C
5.6
-
L-idose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
6.6
-
L-Lyxose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
18.4
-
L-Lyxose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.89
-
NAD+
-
pH 7, 25C
0.92
-
NAD+
-
pH 7.0, 25C
4.7
-
NADH
P31867
mutant K21A, pH 6.5, 35C
12
-
NADH
-
pH 6.0, mutant enzyme K270S/N272P/S271G/R276F
12.7
-
NADH
P31867
mutant K21A/N272D, pH 6.5, 35C
13.1
-
NADH
P31867
wild-type, pH 6.5, 35C
14.2
-
NADH
-
pH 7.0, 25C
15.4
-
NADH
-
pH 6.0, wild-type enzyme, wild-type enzyme
18.1
-
NADH
-
pH 7, 25C
310
-
NADH
-
pH 6.3, 25C
15750
-
NADH
C5J3R6
wild-type, pH 6.5, 37C
25110
-
NADH
C5J3R6
mutant K271R/N273, pH 6.5, 37C
0.82
-
NADP+
-
pH 7, 25C
2.6
-
NADPH
-
pH 6.0, mutant enzyme K270S/N272P/S271G/R276F
21.9
-
NADPH
-
pH 7, 25C
27.5
-
NADPH
-
pH 6.0, wild-type enzyme, wild-type enzyme
40.27
-
NADPH
P31867
wild-type, pH 6.5, 35C
3600
-
NADPH
-
pH 6.3, 25C
100900
-
NADPH
C5J3R6
mutant K271R/N273, pH 6.5, 37C
324000
-
NADPH
C5J3R6
wild-type, pH 6.5, 37C
5.9
-
Pentanal
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
20.7
-
Pentanal
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
4.6
-
propionaldehyde
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
6.9
-
propionaldehyde
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
0.87
-
xylitol
-
pH 7, 25C
0.92
-
xylitol
-
pH 7.0, 25C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
3.7
-
2-deoxy-D-galactose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
2412
6.3
-
2-deoxy-D-galactose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
2412
9.3
-
2-deoxy-D-galactose
-
pH 7.0, 25C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
2412
17
-
2-deoxy-D-galactose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
2412
28
-
2-deoxy-D-galactose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
2412
38
-
2-deoxy-D-galactose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
2412
117
-
2-deoxy-D-glucose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
2416
36.8
-
2-deoxy-D-ribose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
2421
20
-
9,10-phenanthrenequinone
-
mutant H113A, pH 7.0, 25C
6039
33
-
9,10-phenanthrenequinone
-
mutant Y51A, pH 7.0, 25C
6039
500
-
9,10-phenanthrenequinone
-
mutant K80A, pH 7.0, 25C
6039
2300
-
9,10-phenanthrenequinone
-
wild-type, pH 7.0, 25C
6039
0.16
-
Butanal
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
8122
0.93
-
Butanal
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
8122
0.028
-
D-arabinose
Q6Y0Z3
pH 6.0
9095
0.089
-
D-Erythrose
Q6Y0Z3
pH 6.0
9131
736
-
D-Erythrose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
9131
1380
-
D-Erythrose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
9131
1457
-
D-fucose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
9148
2070
-
D-fucose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
9148
0.16
-
D-galactose
-
pH 6.3, 25C
9162
5.5
-
D-galactose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
9162
9
-
D-galactose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
9162
70
-
D-galactose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
9162
249
-
D-galactose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
9162
265
-
D-galactose
-
pH 7.0, 25C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
9162
627
-
D-galactose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
9162
0.05
-
D-glucose
-
pH 6.3, 25C
9202
8.3
-
D-glucose
-
pH 7.0, 25C, 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, 25C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
9202
170
-
D-glucose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
9202
188
-
D-glucose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
9202
833
-
D-glucose
-
pH 7.0, 25C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
9202
1370
-
D-glucose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
9202
0.043
-
D-ribose
Q6Y0Z3
pH 6.0
9348
0.75
-
D-ribose
-
pH 6.3, 25C
9348
72
-
D-ribose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
9348
134
-
D-ribose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
9348
0.0009
-
D-xylose
-
mutant K80A, pH 7.0, 25C
9403
0.019
-
D-xylose
Q6Y0Z3
pH 6.0, coenzyme: NADPH
9403
0.18
-
D-xylose
-
pH 7.0, 25C
9403
0.2
-
D-xylose
O74237
pH 6.0, 25C, cosubstrate: NADH
9403
0.2
-
D-xylose
-
mutant H113A, pH 7.0, 25C
9403
0.21
-
D-xylose
-
pH 7, 25C, cosubstrate: NADH
9403
0.296
-
D-xylose
-
pH 7, 25C, cosubstrate: NADPH
9403
0.89
-
D-xylose
-
pH 7.0, 22C, wild-type enzyme
9403
1.5
-
D-xylose
Q6Y0Z3
pH 6.0, coenzyme: NADH
9403
1.8
-
D-xylose
-
pH 6.3, 25C, cosubstrate: NADPH
9403
6.2
-
D-xylose
-
pH 6.0, cofactor: NADPH, mutant enzyme K270S/N272P/S271G/R276F
9403
6.5
-
D-xylose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme W23Y, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
9403
9
-
D-xylose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
9403
14
-
D-xylose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
9403
19
-
D-xylose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
9403
25
-
D-xylose
-
pH 7.0, 25C, cofactor: NADH, mutant enzyme W23F, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
9403
81.7
-
D-xylose
-
pH 6.0, cofactor: NADH, mutant enzyme K270S/N272P/S271G/R276F
9403
680
-
D-xylose
-
pH 7.0, 25C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose
9403
1460
-
D-xylose
-
pH 6.0, cofactor: NADH, wild-type enzyme
9403
1470
-
D-xylose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
9403
1690
-
D-xylose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
9403
4648
-
D-xylose
-
pH 6.0, cofactor: NADPH, wild-type enzyme
9403
27430
-
D-xylose
A9QVV8
pH 5.5, 45C
9403
0.3
-
DL-glyceraldehyde
-
pH 7.0, 25C, 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, 25C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
10100
5.8
-
DL-glyceraldehyde
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
10100
8.9
-
DL-glyceraldehyde
-
pH 7.0, 25C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose
10100
21.3
-
DL-glyceraldehyde
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
10100
0.75
-
L-arabinose
-
pH 6.3, 25C
12082
482
-
L-arabinose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
12082
1230
-
L-arabinose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
12082
46
-
L-Lyxose
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
12318
157
-
L-Lyxose
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
12318
15.2
-
NAD+
-
pH 7, 25C
14330
34.1
-
NAD+
-
pH 7.0, 25C
14330
81.7
-
NADH
-
pH 6.0, mutant enzyme K270S/N272P/S271G/R276F
14331
146.8
-
NADH
P31867
mutant K21A, pH 6.5, 35C
14331
614
-
NADH
P31867
wild-type, pH 6.5, 35C
14331
713
-
NADH
-
pH 7, 25C
14331
946.7
-
NADH
-
pH 7.0, 25C
14331
1271
-
NADH
P31867
mutant K21A/N272D, pH 6.5, 35C
14331
1460
-
NADH
-
pH 6.0, wild-type enzyme, wild-type enzyme
14331
13900
-
NADH
Q6Y0Z3
pH 6.0
14331
59920
-
NADH
C5J3R6
pH 6.5, 37C, wild-type enzyme; wild-type, pH 6.5, 37C
14331
83750
-
NADH
C5J3R6
mutant K271R/N273, pH 6.5, 37C; pH 6.5, 37C, mutant enzyme K271R/N273D
14331
30.9
-
NADP+
-
pH 7, 25C
27497
6.2
-
NADPH
-
pH 6.0, mutant enzyme K270S/N272P/S271G/R276F
27498
33.3
-
NADPH
-
pH 6.3, 25C
27498
127
-
NADPH
Q6Y0Z3
pH 6.0
27498
1592
-
NADPH
P31867
wild-type, pH 6.5, 35C
27498
4610
-
NADPH
-
pH 7, 25C
27498
4648
-
NADPH
-
pH 6.0, wild-type enzyme, wild-type enzyme
27498
135500
-
NADPH
C5J3R6
mutant K271R/N273, pH 6.5, 37C; pH 6.5, 37C, mutant enzyme K271R/N273D
27498
1466000
-
NADPH
C5J3R6
pH 6.5, 37C, wild-type enzyme; wild-type, pH 6.5, 37C
27498
0.4
-
Pentanal
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
15266
5.31
-
Pentanal
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
15266
0.035
-
propionaldehyde
-
pH 7.0, 25C, dual specific xylose reductase, cofactor: NADH
15841
0.09
-
propionaldehyde
-
pH 7.0, 25C, NADPH-dependent monospecific xylose reductase, cofactor: NADPH
15841
0.0034
-
xylitol
-
pH 7, 25C
17917
0.0044
-
xylitol
-
pH 7.0, 25C
17917
0.031
-
xylitol
Q6Y0Z3
pH 6.0
17917
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0239
-
ATP
-
pH 7, 25C, variable substrate: NADH
0.034
-
Cu2+
Q6Y0Z3
pH 6.0
0.074
-
NAD+
-
pH 7.0, 25C, dual specific xylose reductase (dsXR)
0.13
-
NAD+
-
concentration of NADH varied
0.18
-
NAD+
Q6Y0Z3
pH 6.0, competitive with NADH
0.195
-
NAD+
-
pH 7.0, 25C
0.325
-
NAD+
Q6Y0Z3
pH 6.0, non-competitive with D-xylose
0.65
-
NAD+
-
concentration of D-xylose varied
0.0019
-
NADH
-
pH 7.0, 25C
0.008
-
NADH
-
pH 7.0, 25C, dual specific xylose reductase (dsXR)
0.016
-
NADH
-
pH 7.0, 25C
0.02
-
NADH
-
pH 7.0, 25C, mutant enzyme D50A
0.02
-
NADH
-
pH 7.0, 25C, monospecific xylose reductase (msXR)
0.0015
-
NADP+
-
pH 7, 25C, variable substrate: NADH
0.006
-
NADP+
-
concentration of NADH varied
0.025
-
NADP+
-
pH 7.0, 25C, monospecific xylose reductase (msXR)
0.03
-
NADP+
-
concentration of NADPH varied
0.053
-
NADP+
-
pH 7.0, 25C, dual specific xylose reductase (dsXR)
0.17
-
NADP+
-
concentration of D-xylose varied
0.014
-
NADPH
-
pH 7.0, 25C, dual specific xylose reductase (dsXR)
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.11
-
Torulopsis molishiama
-
-
0.16
0.4
-
recombinant enzyme in transgenic strains of Saccharomyces cerevisiae
0.48
-
-
cell extract
1.39
1.56
-
NADH-dependent activity
2
-
-
mutant enzyme Y49F
2.4
-
P31867
mutant K21A, substrate NADH, pH 6.5, 35C
2.86
-
-
recombinant strain LNG2, pH 7.0, 25C, xylose reduction with NADPH
3.4
-
-
NADPH-dependent activity
4.84
-
-
NADPH-dependent activity
4.93
-
-
NADH-dependent activity
5
-
P31867
mutant K21A, substrate NADPH, pH 6.5, 35C
6.6
-
P31867
wild-type, substrate NADH, pH 6.5, 35C
10.37
-
-
NADH-dependent activity
16.7
-
-
NADH-dependent activity
20.3
-
P31867
wild-type, substrate NADPH, pH 6.5, 35C
20.64
-
-
-
22.1
-
-
NADPH-dependent activity
23.2
-
-
NADPH-dependent activity
41.7
-
Q6Y0Z3
-
47.8
-
-
D-xylose reductase 3
56.9
-
-
D-xylose reductase 1
81
-
-
D-xylose reductase 2
104
-
-
wild-type enzyme
251.5
-
A9QVV8
cofactor: NADPH
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
-
A9QVV8
-
6
-
-
xylose reduction
6
-
Q6Y0Z3
reduction of D-xylose
6
-
-
NADH- and NADPH-dependent activity
6
-
-
D-xylose reductase 1, D-xylose reductase 2
6.3
-
-
assay at
6.5
-
A0MTG4
with NADH as cofactor
6.5
-
C5J3R6
; wild-type and double-mutant (K271R/N273D) protein
7
-
-
assay at
7
-
-
assay at
7
-
-
assay at
7.2
-
F2YCN5
assay at
8.9
-
-
xylitol oxidation
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
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
6.5
A9QVV8
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
Q6Y0Z3
pH 5.0: 81% of maximal activity, pH 7.0: about 65% of maximal activity
5
8
-
the ratio of activities with NADH and NADPH is approximately constant between pH 5 and 8
5
8
C5J3R6
; active from pH 5 to pH 8, wild-type and double-mutant (K271R/N273D) protein
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
assay at
25
-
-
assay at
25
-
-
assay at
25
-
-
assay at
30
-
-
assay at
35
40
A0MTG4
-
37
-
Q6Y0Z3
assay at
37
-
C5J3R6
; wild-type and double-mutant (K271R/N273D) protein
45
-
A9QVV8
-
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
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
40
-
activity increases linearly from 25C to 50C
25
65
-
25C: about 55% of maximal activity, 65C: about 65% of maximal activity
30
60
C5J3R6
active from 30C to 60C, wild-type and double-mutant (K271R/N273D) protein
30
60
A9QVV8
30C: about 70% of maximal activity, 60C: about 45% of maximal activity
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.1
-
-
D-xylose reductase 2, isoelectric focusing, pH-range: 2.5-5.0
4.15
-
-
D-xylose reductase 1, isoelectric focusing, pH-range: 2.5-5.0
4.7
-
-
isoelectric focusing
5.1
-
C1K8Y9
calculated
5.19
-
Q6Y0Z3
calculated from sequence
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Candida diddensiae F-3, Candida shehatae Y-1632, Candida silvanorum VGI-II, Candida tropicalis Y-456, Kluyveromyces marxianus Y-488, Meyerozyma guilliermondii Y-1017, Scheffersomyces stipitis Y-2160, Torulopsis molishiama 55
-
microaerobically grown
-
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
48000
-
-
gel filtration
53000
-
-
gel filtration
58000
-
-
D-xylose reductase 1, D-xylose reductase 2, D-xylose reductase 3, gel filtration
60000
-
-
gel filtration
63000
65000
-
gel filtration
69000
-
Q6Y0Z3
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
C1K8Y9
x * 36600, calculated; x * 37000, SDS-PAGE
?
C5J3R6
x * 36000, calculated, x * 40000, SDS-PAGE; x * 40000, wild-type and double-mutant (K271R/N273D) histidine-tagged protein, SDS-PAGE
?
A9QVV8
x * 37100, SDS-PAGE
?
-
x * 37000, wild-type and mutant enzyme Y49F, SDS-PAGE
?
P31867
x * 36000, SDS-PAGE of recombinant enzyme
?
Candida tropicalis SCTCC 300249
-
x * 37100, SDS-PAGE
-
dimer
Q6Y0Z3
2 * 36400, SDS-PAGE; 2 * 36629, calculated from sequence
dimer
-
2 * 38400, SDS-PAGE
dimer
-
2 * 34000, SDS-PAGE
dimer
-
2 * 29000, SDS-PAGE
dimer
-
2 * 364978, D-xylose reductase 1, ion-spray mass spectrometry; 2 * 365540, D-xylose reductase 2, ion-spray mass spectrometry
dimer
Candida parapsilosis KFCC-10875
-
2 * 36400, SDS-PAGE; 2 * 36629, calculated from sequence
-
dimer
Candida tropicalis IF0 0618
-
2 * 364978, D-xylose reductase 1, ion-spray mass spectrometry; 2 * 365540, D-xylose reductase 2, ion-spray mass spectrometry
-
monomer
-
1 * 43000, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the purified N309D mutant is crystallized by the hanging-drop vapour-diffusion method at 25C. 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
C1K8Y9
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
8
-
below pH 5 and above pH 8.0 the enzyme is inactivated within 3-6 days
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
-
Q6Y0Z3
60 days, 50% loss of activity
20
-
Q6Y0Z3
8 days, 50% loss of activity
21
-
-
at room temperature stable for more than 1 month
25
-
-
half-life: more than 2 months
30
35
-
48 h, stability starts to decrease above 30-35C
30
-
Q6Y0Z3
3 days, 50% loss of activity
40
-
-
half-life: 94 min
45
-
Q6Y0Z3
4.5 h, 50% loss of activity
50
-
Q6Y0Z3
2 min, 50% loss of activity
60
-
C5J3R6
retained 28% of activity at 60C
60
-
-
1 h, D-xylose reductase 1, D-xylose reductase 2, complete loss of activity
60
-
A9QVV8
1 h, 80% loss of activity, half-life is around 15 min
additional information
-
-
non-ionic detergents and bovine serum albumin stabilize the enzyme to a significant extent during long-term incubation at 25C, 30C or 38C
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
stable enzyme at 25C in phosphate and Tris buffer of various ionic strengths between pH 6.0 and 7.0
-
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the enzyme undergoes thiol oxidation during storage or purification
-
286260
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-18C, 4 months, activity in cell extract remains stable
-
38C, 3 h, activity in cell extract remains stable
-
4C, 3 h, activity in cell extract remains stable
-
4C, enzyme retains activity for several months
A0MTG4
-20C, pure enzyme preparation is stable for more than 4 months
-
4C, pure enzyme preparation is stable for more than 4 months
-
4C, stable for several months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
; recombinant enzyme
C1K8Y9
recombinant enzyme
A9QVV8
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
-
wild-type and mutant enzyme Y49F
-
recombinant enzyme
P31867
; recombinant protein
C5J3R6
recombinant His-tagged enzyme from Escherichia coli strain UT5600 by nickel affinity chromatography
F2YCN5
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Candida tropicalis increases production of ethanol and glycerol
Q6Y0Z3
expression in Escherichia coli as a His6-tagged fusion protein
A0MTG4
expression in Escherichia coli
O74237
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
-
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
-
expressed in Escherichia coli BL21 (DE3), subloned into the pYES2 vector and transformed into Saccharomyces cerevisiae W303-1A
A9QVV8
expression in Escherichia coli; subcloned into the expression plasmid pET28a(+) and subsequently transformed into Escherichia coli BL21(DE3)pLysS cells for recombinant protein expression
C1K8Y9
construction of Kluyveromyces marxianus strains expressing Pichia stipitis Psxyl1 genes leading to reversed cofactor specificity and ethanol and xylitol production
-
expression in Escherichia coli as a His6-tagged fusion protein in high yield
-
expression in Escherichia coli
-
expression in Escherichia coli
P31867
expression of His-tagged enzyme in Escherichia coli
-
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
-
expression in Escherichia coli; expression of histidine-tagged wild type Texr and mutant Texr K271R/N273D in Escherichia coli BL21-Star DE3
C5J3R6
gene ZMO0976, expression of His-tagged enzyme in Escherichia coli strain UT5600
F2YCN5
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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
-, Q9P8R5
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
Candida tropicalis ATCC 20336
-
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
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
N309A
-
the 30fold preference of the wild-type for D-galactose compared with 2-deoxy-D-galactose is lost completely in the mutant. Replacement of Asn309 with alanine or aspartic acid disrupts the function of the original side chain in donating a hydrogen atom for bonding with the substrate C-2(R) hydroxy group, thus causing a loss of transition-state stabilization energy of 89 kJ/mol
N309D
-
the 30fold preference of the wild-type for D-galactose compared with 2-deoxy-D-galactose is lost completely in the mutant. Comparison of the 2.4 A X-ray crystal structure of mutant N309D bound to NAD+ with the previous structure of the wild-type holoenzyme reveals no major structural perturbations. Replacement of Asn309 with alanine or aspartic acid disrupts the function of the original side chain in donating a hydrogen atom for bonding with the substrate C-2(R) hydroxy group, thus causing a loss of transition-state stabilization energy of 89 kJ/mol
W23F
-
mutant catalyses NADH-dependent reduction of xylose with 4% of the wild-type efficiency (kcat/Km), but improves the wild-type selectivity for utilization of ketones, relative to xylose, by factors of 156
W23Y
-
mutant catalyses NADH-dependent reduction of xylose with 1% of the wild-type efficiency (kcat/Km), but improves the wild-type selectivity for utilization of ketones, relative to xylose, by factors of 471
K274M
-
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
Y49F
-
more than 98% loss of activity compared to wild-type enzyme
N272D
Saccharomyces cerevisiae TMB 3420
-
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
Saccharomyces cerevisiae TMB 3420
-
site-directed mutagenesis, results in strain TMB 3421, the mutations enable the yeast for anaerobic growth on xylose displayed higher aerobic growth rates
-
P275Q
Saccharomyces cerevisiae TMB 3420
-
site-directed mutagenesis, results in strain TMB 3423, the mutation enables the yeast for anaerobic growth on xylose displayed higher aerobic growth rates
-
K21A
P31867
strong preference for NADH over NADPH
K21A/N272D
P31867
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
K271R/N273D
C5J3R6
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
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
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
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
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
Candida tropicalis ATCC 20336
-
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
Corynebacterium glutamicum ATCC13032
-
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
Kluyveromyces marxianus YHJ010
-
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
-
K74M/N276D
-
site-directed mutagenesis, the mutant enzyme shows increased activity and altered kinetics compared to the wild-type enzyme
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
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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 8C 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
O74237
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
synthesis
-
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
O74237
production of xylitol
synthesis
-
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
Candida tenuis CBS 4435
-
production of xylitol
-
synthesis
-
the enzyme is useful for xylitol bioproduction, profiles, overview
synthesis
Meyerozyma guilliermondii FTI, Meyerozyma guilliermondii FTI 20037
-
the enzyme is useful for xylitol bioproduction, profiles, overview
-
synthesis
-
this enzyme is one of the most active xylose reductases and may be used for the in vitro production of xylitol
synthesis
-
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
-
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