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Literature summary extracted from

  • Mouro, A.; Santos, A.; Agnolo, D.; Gubert, G.; Bon, E.; Rosa, C.; Fonseca, C.; Stambuk, B.
    Combining xylose reductase from Spathaspora arborariae with xylitol dehydrogenase from Spathaspora passalidarum to promote xylose consumption and fermentation into xylitol by Saccharomyces cerevisiae (2020), Fermentation, 6, 72 .
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.9 gene SpXYL2.2, recombinant expression of the enzyme in Saccharomyces cerevisiae strain CEN.PK2-1C (MATa leu2-3112 ura3-52 trp1-289 his3-DELTA1 MAL2-8c SUC2), subcloning in Escherichia coli strain DH5alpha, co-expression with xylose reductase from Spathaspora arborariae Spathaspora passalidarum
1.1.1.307 gene SpXYL1.1, recombinant expression of the enzyme in Saccharomyces cerevisiae strain CEN.PK2-1C (MATa leu2-3112 ura3-52 trp1-289 his3-DELTA1 MAL2-8c SUC2), subcloning in Escherichia coli strain DH5alpha, co-expression with xylitol dehydrogenase from Spathaspora passalidarum strain UFMG-CM-Y474 Spathaspora arborariae

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.9 additional information combination of xylose reductase from Spathaspora arborariae with xylitol dehydrogenase from Spathaspora passalidarum to promote xylose consumption and fermentation into xylitol in Saccharomyces cerevisiae. Recombinant co-expression of Spathaspora arborariae xylose reductase gene (SaXYL1) that accepts both NADH and NADPH as co-substrates, and of Spathaspora passalidarum strain UFMG-HM.19.1AT NADPH-dependent xylose reductase (SpXYL1.1 gene) or the SpXYL2.2 gene from Spathaspora passalidarum strain UFMG-CM-Y474 in a Saccharomyces cerevisiae strain overexpressing the native XKS1 gene encoding xylulokinase, as well as being deleted in the alkaline phosphatase encoded by the PHO13 gene. Strains expressing the Spathaspora enzymes consumes xylose with xylitol as the major fermentation product. Higher specific growth rates, xylose consumption, and xylitol volumetric productivities are obtained by the co-expression of the SaXYL1 and SpXYL2.2 genes, when compared with the co-expression of the NADPH-dependent SpXYL1.1 xylose reductase. During glucose-xylose co-fermentation by the strain with co-expression of the SaXYL1 and SpXYL2.2 genes, both ethanol and xylitol are produced efficiently. Method development and evaluation, detailed overview Spathaspora passalidarum
1.1.1.307 additional information combination of xylose reductase from Spathaspora arborariae with xylitol dehydrogenase from Spathaspora passalidarum to promote xylose consumption and fermentation into xylitol in Saccharomyces cerevisiae. Recombinant co-expression of Spathaspora arborariae xylose reductase gene (SaXYL1) that accepts both NADH and NADPH as co-substrates, and of Spathaspora arborariae strain UFMG-HM.19.1AT NADPH-dependent xylose reductase (SpXYL1.1 gene) or the SpXYL2.2 gene from Spathaspora passalidarum strain UFMG-CM-Y474 in a Saccharomyces cerevisiae strain overexpressing the native XKS1 gene encoding xylulokinase, as well as being deleted in the alkaline phosphatase encoded by the PHO13 gene. Strains expressing the Spathaspora enzymes consumes xylose with xylitol as the major fermentation product. Higher specific growth rates, xylose consumption, and xylitol volumetric productivities are obtained by the co-expression of the SaXYL1 and SpXYL2.2 genes, when compared with the co-expression of the NADPH-dependent SpXYL1.1 xylose reductase. During glucose-xylose co-fermentation by the strain with co-expression of the SaXYL1 and SpXYL2.2 genes, both ethanol and xylitol are produced efficiently. Method development and evaluation, detailed overview Spathaspora arborariae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.9 additional information
-
 additional information Michaelis-Menten kinetics Spathaspora passalidarum
1.1.1.9 0.52
-
 NAD+ recombinant enzyme, pH 9.0, 35°C Spathaspora passalidarum
1.1.1.9 86
-
 xylitol recombinant enzyme, pH 9.0, 35°C Spathaspora passalidarum
1.1.1.307 0.0659
-
 NADPH recombinant enzyme, pH 6.0, 30°C Spathaspora arborariae
1.1.1.307 53.3
-
 D-xylose recombinant enzyme, pH 6.0, 30°C Spathaspora arborariae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.9 xylitol + NAD+ Spathaspora passalidarum
-
 D-xylulose + NADH + H+
-
 r
1.1.1.9 xylitol + NAD+ Spathaspora passalidarum UFMG-CM-Y474
-
 D-xylulose + NADH + H+
-
 r
1.1.1.307 D-xylose + NADH + H+ Spathaspora arborariae
-
 xylitol + NAD+
-
 r
1.1.1.307 D-xylose + NADH + H+ Spathaspora arborariae UFMG-HM.19.1AT
-
 xylitol + NAD+
-
 r
1.1.1.307 D-xylose + NADPH + H+ Spathaspora arborariae
-
 xylitol + NADP+
-
 r
1.1.1.307 D-xylose + NADPH + H+ Spathaspora arborariae UFMG-HM.19.1AT
-
 xylitol + NADP+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.9 Spathaspora passalidarum
-
 isolated from rotting wood in Roraima, Brazil
-
1.1.1.9 Spathaspora passalidarum UFMG-CM-Y474
-
 isolated from rotting wood in Roraima, Brazil
-
1.1.1.307 Spathaspora arborariae
-
 isolated from rotting wood in Minas Gerais, Brazil
-
1.1.1.307 Spathaspora arborariae UFMG-HM.19.1AT
-
 isolated from rotting wood in Minas Gerais, Brazil
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.9 0.003
-
 native enzyme, substrate xylose and NADP+, pH 9.0, 35°C Spathaspora passalidarum
1.1.1.9 1.29
-
 native enzyme, substrate xylitol and NAD+, pH 9.0, 35°C Spathaspora passalidarum
1.1.1.9 2.2
-
 recombinant enzyme expressed in Saccharomyces cerevisiae strain CEN.PK2-1C, substrate xylitol and NAD+, pH 9.0, 35°C Spathaspora passalidarum
1.1.1.9 2.95
-
 recombinant enzyme expressed in Saccharomyces cerevisiae strain CEN.PK2-1C, co-expressed with xylose reductase from Spathaspora arborariae, substrate xylitol and NAD+, pH 9.0, 35°C Spathaspora passalidarum
1.1.1.307 0.118
-
 native enzyme, substrate xylose and NADH, pH 6.0, 30°C Spathaspora arborariae
1.1.1.307 0.455
-
 native enzyme, substrate xylose and NADPH, pH 6.0, 30°C Spathaspora arborariae
1.1.1.307 5.12
-
 recombinant enzyme expressed in Saccharomyces cerevisiae strain CEN.PK2-1C, co-expressed with xylitol dehydrogenase from Spathaspora passalidarum strain UFMG-CM-Y474, substrate xylose and NADH, pH 6.0, 30°C Spathaspora arborariae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.9 xylitol + NAD+
-
 Spathaspora passalidarum D-xylulose + NADH + H+
-
 r
1.1.1.9 xylitol + NAD+
-
 Spathaspora passalidarum UFMG-CM-Y474 D-xylulose + NADH + H+
-
 r
1.1.1.307 D-xylose + NADH + H+
-
 Spathaspora arborariae xylitol + NAD+
-
 r
1.1.1.307 D-xylose + NADH + H+ about 25% of the activity with NADPH Spathaspora arborariae xylitol + NAD+
-
 r
1.1.1.307 D-xylose + NADH + H+
-
 Spathaspora arborariae UFMG-HM.19.1AT xylitol + NAD+
-
 r
1.1.1.307 D-xylose + NADH + H+ about 25% of the activity with NADPH Spathaspora arborariae UFMG-HM.19.1AT xylitol + NAD+
-
 r
1.1.1.307 D-xylose + NADPH + H+
-
 Spathaspora arborariae xylitol + NADP+
-
 r
1.1.1.307 D-xylose + NADPH + H+ NADPH is the preferred cofactor Spathaspora arborariae xylitol + NADP+
-
 r
1.1.1.307 D-xylose + NADPH + H+
-
 Spathaspora arborariae UFMG-HM.19.1AT xylitol + NADP+
-
 r
1.1.1.307 D-xylose + NADPH + H+ NADPH is the preferred cofactor Spathaspora arborariae UFMG-HM.19.1AT xylitol + NADP+
-
 r

Synonyms

EC Number Synonyms Comment Organism
1.1.1.9 SpXYL2.2
-
 Spathaspora passalidarum
1.1.1.9 XDH
-
 Spathaspora passalidarum
1.1.1.9 xylitol dehydrogenase
-
 Spathaspora passalidarum
1.1.1.307 SaXYL1
-
 Spathaspora arborariae
1.1.1.307 SpXYL1.1
-
 Spathaspora arborariae
1.1.1.307 xylose reductase
-
 Spathaspora arborariae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.9 35
-
 xylitol oxidation, assay at Spathaspora passalidarum
1.1.1.307 30
-
 assay at Spathaspora arborariae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.9 9
-
 xylitol oxidation, assay at Spathaspora passalidarum
1.1.1.307 6
-
 assay at Spathaspora arborariae

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.9 NAD+
-
 Spathaspora passalidarum
1.1.1.9 NADH
-
 Spathaspora passalidarum
1.1.1.307 additional information Spathaspora arborariae xylose reductase accepts both NADH and NADPH as co-substrates, gene SpXYL1.1 encodes for a xylose reductase with higher NADH activity compared to other XRs. The NADH-dependent of activity of Xyl1 is 25% compared to NADPH Spathaspora arborariae
1.1.1.307 NAD+
-
 Spathaspora arborariae
1.1.1.307 NADH
-
 Spathaspora arborariae
1.1.1.307 NADP+
-
 Spathaspora arborariae
1.1.1.307 NADPH
-
 Spathaspora arborariae