Cloned (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
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] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0659 | - |
NADPH | recombinant enzyme, pH 6.0, 30°C | Spathaspora arborariae | |
53.3 | - |
D-xylose | recombinant enzyme, pH 6.0, 30°C | Spathaspora arborariae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-xylose + NADH + H+ | Spathaspora arborariae | - |
xylitol + NAD+ | - |
r | |
D-xylose + NADH + H+ | Spathaspora arborariae UFMG-HM.19.1AT | - |
xylitol + NAD+ | - |
r | |
D-xylose + NADPH + H+ | Spathaspora arborariae | - |
xylitol + NADP+ | - |
r | |
D-xylose + NADPH + H+ | Spathaspora arborariae UFMG-HM.19.1AT | - |
xylitol + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Spathaspora arborariae | - |
isolated from rotting wood in Minas Gerais, Brazil | - |
Spathaspora arborariae UFMG-HM.19.1AT | - |
isolated from rotting wood in Minas Gerais, Brazil | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.118 | - |
native enzyme, substrate xylose and NADH, pH 6.0, 30°C | Spathaspora arborariae |
0.455 | - |
native enzyme, substrate xylose and NADPH, pH 6.0, 30°C | Spathaspora arborariae |
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-xylose + NADH + H+ | - |
Spathaspora arborariae | xylitol + NAD+ | - |
r | |
D-xylose + NADH + H+ | about 25% of the activity with NADPH | Spathaspora arborariae | xylitol + NAD+ | - |
r | |
D-xylose + NADH + H+ | - |
Spathaspora arborariae UFMG-HM.19.1AT | xylitol + NAD+ | - |
r | |
D-xylose + NADH + H+ | about 25% of the activity with NADPH | Spathaspora arborariae UFMG-HM.19.1AT | xylitol + NAD+ | - |
r | |
D-xylose + NADPH + H+ | - |
Spathaspora arborariae | xylitol + NADP+ | - |
r | |
D-xylose + NADPH + H+ | NADPH is the preferred cofactor | Spathaspora arborariae | xylitol + NADP+ | - |
r | |
D-xylose + NADPH + H+ | - |
Spathaspora arborariae UFMG-HM.19.1AT | xylitol + NADP+ | - |
r | |
D-xylose + NADPH + H+ | NADPH is the preferred cofactor | Spathaspora arborariae UFMG-HM.19.1AT | xylitol + NADP+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
SaXYL1 | - |
Spathaspora arborariae |
SpXYL1.1 | - |
Spathaspora arborariae |
xylose reductase | - |
Spathaspora arborariae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Spathaspora arborariae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Spathaspora arborariae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
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 | |
NAD+ | - |
Spathaspora arborariae | |
NADH | - |
Spathaspora arborariae | |
NADP+ | - |
Spathaspora arborariae | |
NADPH | - |
Spathaspora arborariae |