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Literature summary for 1.1.1.307 extracted from

  • Quehenberger, J.; Reichenbach, T.; Baumann, N.; Rettenbacher, L.; Divne, C.; Spadiut, O.
    Kinetics and predicted structure of a novel xylose reductase from Chaetomium thermophilum (2019), Int. J. Mol. Sci., 20, 185 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.1.1.307

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of C-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3) Thermochaetoides thermophila

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics Thermochaetoides thermophila
0.119
-
 NADH recombinant enzyme, pH 6.5, 30°C, with D-xylose Thermochaetoides thermophila
0.135
-
 NADPH recombinant enzyme, pH 6.5, 30°C, with D-xylose Thermochaetoides thermophila
22.3
-
 D-xylose recombinant enzyme, pH 6.5, 30°C Thermochaetoides thermophila
25.4
-
 D-xylose recombinant enzyme, pH 6.5, 55°C Thermochaetoides thermophila
62.5
-
 L-arabinose recombinant enzyme, pH 6.5, 30°C, with NADPH Thermochaetoides thermophila
241
-
 D-galactose recombinant enzyme, pH 6.5, 30°C, with NADPH Thermochaetoides thermophila
471
-
 D-glucose recombinant enzyme, pH 6.5, 30°C, with NADPH Thermochaetoides thermophila

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
 Thermochaetoides thermophila 5829
-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-xylose + NADPH + H+ Thermochaetoides thermophila
-
 xylitol + NADP+
-
 r
D-xylose + NADPH + H+ Thermochaetoides thermophila IMI 039719
-
 xylitol + NADP+
-
 r
D-xylose + NADPH + H+ Thermochaetoides thermophila DSM 1495
-
 xylitol + NADP+
-
 r
D-xylose + NADPH + H+ Thermochaetoides thermophila CBS 144.50
-
 xylitol + NADP+
-
 r

Organism

Organism UniProt Comment Textmining
Thermochaetoides thermophila G0SCE7
-
-
Thermochaetoides thermophila CBS 144.50 G0SCE7
-
-
Thermochaetoides thermophila DSM 1495 G0SCE7
-
-
Thermochaetoides thermophila IMI 039719 G0SCE7
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme 25.5fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration to near homogeneity Thermochaetoides thermophila

Source Tissue

Source Tissue Comment Organism Textmining
additional information optimal growth temperature of the organism is 50-55°C Thermochaetoides thermophila
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-galactose + NADPH + H+
-
 Thermochaetoides thermophila galactitol + NADP+
-
 r
D-galactose + NADPH + H+
-
 Thermochaetoides thermophila IMI 039719 galactitol + NADP+
-
 r
D-galactose + NADPH + H+
-
 Thermochaetoides thermophila DSM 1495 galactitol + NADP+
-
 r
D-galactose + NADPH + H+
-
 Thermochaetoides thermophila CBS 144.50 galactitol + NADP+
-
 r
D-glucose + NADPH + H+
-
 Thermochaetoides thermophila glucitol + NADP+
-
 r
D-ribose + NADPH + H+
-
 Thermochaetoides thermophila ribitol + NADP+
-
 r
D-xylose + NADH + H+
-
 Thermochaetoides thermophila xylitol + NAD+
-
 r
D-xylose + NADH + H+
-
 Thermochaetoides thermophila IMI 039719 xylitol + NAD+
-
 r
D-xylose + NADH + H+
-
 Thermochaetoides thermophila DSM 1495 xylitol + NAD+
-
 r
D-xylose + NADH + H+
-
 Thermochaetoides thermophila CBS 144.50 xylitol + NAD+
-
 r
D-xylose + NADPH + H+
-
 Thermochaetoides thermophila xylitol + NADP+
-
 r
D-xylose + NADPH + H+ best substrates Thermochaetoides thermophila xylitol + NADP+
-
 r
D-xylose + NADPH + H+
-
 Thermochaetoides thermophila IMI 039719 xylitol + NADP+
-
 r
D-xylose + NADPH + H+ best substrates Thermochaetoides thermophila IMI 039719 xylitol + NADP+
-
 r
D-xylose + NADPH + H+
-
 Thermochaetoides thermophila DSM 1495 xylitol + NADP+
-
 r
D-xylose + NADPH + H+ best substrates Thermochaetoides thermophila DSM 1495 xylitol + NADP+
-
 r
D-xylose + NADPH + H+
-
 Thermochaetoides thermophila CBS 144.50 xylitol + NADP+
-
 r
D-xylose + NADPH + H+ best substrates Thermochaetoides thermophila CBS 144.50 xylitol + NADP+
-
 r
L-arabinose + NADPH + H+
-
 Thermochaetoides thermophila arabitol + NADP+
-
 r
additional information the enzyme exhibits dual cofactor specificity for NADPH and NADH and prefers D-xylose over other pentoses and investigated hexoses. No or poor activity with D-lyxose, D-mannose, L-xylose, and D-arabinose. L-arabinose, being structurally similar to D-xylose, has a kcat/KM value of still 33% of the value for D-xylose, whereas D-galactose and D-glucose result in only 6.3% and 1.3% of the kcat/KM for D-xylose. The difference in the catalytic efficiency is mainly a result from different KM values rather than differences in kcat Thermochaetoides thermophila ?
-
-
additional information the enzyme exhibits dual cofactor specificity for NADPH and NADH and prefers D-xylose over other pentoses and investigated hexoses. No or poor activity with D-lyxose, D-mannose, L-xylose, and D-arabinose. L-arabinose, being structurally similar to D-xylose, has a kcat/KM value of still 33% of the value for D-xylose, whereas D-galactose and D-glucose result in only 6.3% and 1.3% of the kcat/KM for D-xylose. The difference in the catalytic efficiency is mainly a result from different KM values rather than differences in kcat Thermochaetoides thermophila IMI 039719 ?
-
-
additional information the enzyme exhibits dual cofactor specificity for NADPH and NADH and prefers D-xylose over other pentoses and investigated hexoses. No or poor activity with D-lyxose, D-mannose, L-xylose, and D-arabinose. L-arabinose, being structurally similar to D-xylose, has a kcat/KM value of still 33% of the value for D-xylose, whereas D-galactose and D-glucose result in only 6.3% and 1.3% of the kcat/KM for D-xylose. The difference in the catalytic efficiency is mainly a result from different KM values rather than differences in kcat Thermochaetoides thermophila DSM 1495 ?
-
-
additional information the enzyme exhibits dual cofactor specificity for NADPH and NADH and prefers D-xylose over other pentoses and investigated hexoses. No or poor activity with D-lyxose, D-mannose, L-xylose, and D-arabinose. L-arabinose, being structurally similar to D-xylose, has a kcat/KM value of still 33% of the value for D-xylose, whereas D-galactose and D-glucose result in only 6.3% and 1.3% of the kcat/KM for D-xylose. The difference in the catalytic efficiency is mainly a result from different KM values rather than differences in kcat Thermochaetoides thermophila CBS 144.50 ?
-
-

Subunits

Subunits Comment Organism
? x * 39244, sequence calculation, x * 41000, recombinant His-tagged enzyme, SDS-PAGE Thermochaetoides thermophila

Synonyms

Synonyms Comment Organism
CTHT_0056950
-
 Thermochaetoides thermophila
CtXR
-
 Thermochaetoides thermophila
NAD(P)H-dependent D-xylose reductase-like protein UniProt Thermochaetoides thermophila
xylose reductase
-
 Thermochaetoides thermophila

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
 higher activity at 55°C compared to 30°C Thermochaetoides thermophila

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30
-
 purified recombinant His-tagged enzyme, no loss of activity after 3 h at 30°C Thermochaetoides thermophila
55 75 purified recombinant His-tagged enzyme, half-lifes are 1.8 min at 55°C, 4.2 s at 65°C, and 2.8 s at 75°C, respectively Thermochaetoides thermophila

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7
-
 D-glucose recombinant enzyme, pH 6.5, 30°C, with NADPH Thermochaetoides thermophila
3.2
-
 NADH recombinant enzyme, pH 6.5, 30°C, with D-xylose Thermochaetoides thermophila
6.5
-
 D-galactose recombinant enzyme, pH 6.5, 30°C, with NADPH Thermochaetoides thermophila
8.8
-
 L-arabinose recombinant enzyme, pH 6.5, 30°C, with NADPH Thermochaetoides thermophila
9.2
-
 D-xylose recombinant enzyme, pH 6.5, 30°C, with NADPH Thermochaetoides thermophila
11.4
-
 NADPH recombinant enzyme, pH 6.5, 30°C, with D-xylose Thermochaetoides thermophila
35.2
-
 D-xylose recombinant enzyme, pH 6.5, 55°C, with NADPH Thermochaetoides thermophila

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 6.5 in 50 mM citrate and 50 mM phosphate buffer Thermochaetoides thermophila
7
-
 in a citrate-disodium phosphate buffer Thermochaetoides thermophila

pH Range

pH Minimum pH Maximum Comment Organism
5 8 over 40% of maximal activity within this range, profile overview Thermochaetoides thermophila

Cofactor

Cofactor Comment Organism Structure
additional information the enzyme exhibits dual cofactor specificity for NADPH and NADH, the catalytic efficiency is significantly higher for NADPH Thermochaetoides thermophila
NADP+
-
 Thermochaetoides thermophila
NADPH modeled cofactor-binding site in CtXR, overview Thermochaetoides thermophila

General Information

General Information Comment Organism
additional information structure homology modeling based on a XR structure from Candida sp. (CaXR) as template (PDB ID 1SM9), analysis of the architecture of the cofactor binding site. Notable CaXR-to-CtXR replacements include N276T, L277R, R280I and Q283S Thermochaetoides thermophila

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.006
-
 D-glucose recombinant enzyme, pH 6.5, 30°C, with NADPH Thermochaetoides thermophila
0.027
-
 D-galactose recombinant enzyme, pH 6.5, 30°C, with NADPH Thermochaetoides thermophila
0.141
-
 L-arabinose recombinant enzyme, pH 6.5, 30°C, with NADPH Thermochaetoides thermophila
0.413
-
 D-xylose recombinant enzyme, pH 6.5, 30°C Thermochaetoides thermophila
1.386
-
 D-xylose recombinant enzyme, pH 6.5, 55°C Thermochaetoides thermophila
26.9
-
 NADH recombinant enzyme, pH 6.5, 30°C, with D-xylose Thermochaetoides thermophila
84.4
-
 NADPH recombinant enzyme, pH 6.5, 30°C, with D-xylose Thermochaetoides thermophila