Literature summary for 1.1.1.431 extracted from

Kratzer, R.; Leitgeb, S.; Wilson, D.K.; Nidetzky, B.
Probing the substrate binding site of Candida tenuis xylose reductase (AKR2B5) with site-directed mutagenesis (2006), Biochem. J., 393, 51-58.

Search for more literature for 1.1.1.431

Crystallization (Commentary)

Crystallization (Comment)	Organism
the purified N309D mutant is crystallized by the hanging-drop vapour-diffusion method at 25°C. The best-diffracting crystals are grown using a well solution consisting of 2.1 M (NH4)2SO4, 100 mM sodium acetate and 100 mM sodium citrate, pH 6.4. Comparison of the 2.4 A X-ray crystal structure of mutant N309D bound to NAD+ with the previous structure of the wild-type holoenzyme reveals no major structural perturbations	Yamadazyma tenuis

Crystallization (Comment)

Organism

the purified N309D mutant is crystallized by the hanging-drop vapour-diffusion method at 25°C. The best-diffracting crystals are grown using a well solution consisting of 2.1 M (NH4)2SO4, 100 mM sodium acetate and 100 mM sodium citrate, pH 6.4. Comparison of the 2.4 A X-ray crystal structure of mutant N309D bound to NAD+ with the previous structure of the wild-type holoenzyme reveals no major structural perturbations

Yamadazyma tenuis

Protein Variants

Protein Variants	Comment	Organism
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	Yamadazyma tenuis
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	Yamadazyma tenuis
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	Yamadazyma tenuis
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	Yamadazyma tenuis
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	Yamadazyma tenuis

Inhibitors

Inhibitors	Comment	Organism	Structure
NADH	-	Yamadazyma tenuis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
2.3	-	NADPH	pH 7.0, 25°C, mutant enzyme D50A	Yamadazyma tenuis
3.2	-	NADPH	pH 7.0, 25°C, wild-type enzyme	Yamadazyma tenuis
38	-	NADH	pH 7.0, 25°C, wild-type enzyme	Yamadazyma tenuis
40	-	NADH	pH 7.0, 25°C, mutant enzyme D50A	Yamadazyma tenuis
334	-	xylitol	pH 7.0, 25°C, wild-type enzyme	Yamadazyma tenuis
537	-	xylitol	pH 7.0, 25°C, mutant enzyme D50A	Yamadazyma tenuis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
D-xylose + NADPH + H+	Yamadazyma tenuis	-	xylitol + NADP+	-	?

Organism

Organism	UniProt	Comment	Textmining
Yamadazyma tenuis	O74237	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Yamadazyma tenuis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2-deoxy-D-galactose + NADH	-	Yamadazyma tenuis	?	-	?
D-galactose + NADH	-	Yamadazyma tenuis	?	-	?
D-glucose + NADH	-	Yamadazyma tenuis	?	-	?
D-glyceraldehyde + NADH	-	Yamadazyma tenuis	?	-	?
D-xylose + NADH + H+	kcat of wilde-type enzyme increases by a factor of 1.73 when NADPH replaces NADH	Yamadazyma tenuis	xylitol + NAD+	-	r
D-xylose + NADPH + H+	-	Yamadazyma tenuis	xylitol + NADP+	-	?
D-xylose + NADPH + H+	kcat of wild-type enzyme increases by a factor of 1.73 when NADPH replaces NADH	Yamadazyma tenuis	xylitol + NADP+	-	r
additional information	in vitro the enzyme also catalyzes the reduction of ketones	Yamadazyma tenuis	?	-	?

Synonyms

Synonyms	Comment	Organism
CtXR	-	Yamadazyma tenuis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Yamadazyma tenuis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Yamadazyma tenuis

Cofactor

Cofactor	Comment	Organism	Structure
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	Yamadazyma tenuis
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	Yamadazyma tenuis

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.0019	-	NADH	pH 7.0, 25°C	Yamadazyma tenuis
0.02	-	NADH	pH 7.0, 25°C, mutant enzyme D50A	Yamadazyma tenuis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.3	-	DL-glyceraldehyde	pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
0.3	-	DL-glyceraldehyde	pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
3.7	-	2-deoxy-D-galactose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
5.5	-	D-galactose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
6.3	-	2-deoxy-D-galactose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
6.5	-	D-xylose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme W23Y, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
8.3	-	D-glucose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
8.3	-	D-glucose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
8.9	-	DL-glyceraldehyde	pH 7.0, 25°C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
9	-	D-galactose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
9	-	D-xylose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309D, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
9.3	-	2-deoxy-D-galactose	pH 7.0, 25°C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
14	-	D-xylose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
17	-	2-deoxy-D-galactose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
19	-	D-xylose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme N309A, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
25	-	D-xylose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme W23F, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
70	-	D-galactose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
188	-	D-glucose	pH 7.0, 25°C, cofactor: NADH, mutant enzyme D50A, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
265	-	D-galactose	pH 7.0, 25°C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
680	-	D-xylose	pH 7.0, 25°C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the 0.02% of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis
833	-	D-glucose	pH 7.0, 25°C, cofactor: NADH, wild-type enzyme, kcat/Km value is corrected for the proportion of open-chain free aldehyde in aqueous solution of xylose	Yamadazyma tenuis