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Literature summary for 1.1.1.9 extracted from

  • Hou, J.; Shen, Y.; Li, X.P.; Bao, X.M.
    Effect of the reversal of coenzyme specificity by expression of mutated Pichia stipitis xylitol dehydrogenase in recombinant Saccharomyces cerevisiae (2007), Lett. Appl. Microbiol., 45, 184-189.
    View publication on PubMed
Search for more literature for 1.1.1.9

Cloned(Commentary)

Cloned (Comment) Organism
genes XYL2 (D207A/I208R/F209S) and XYL2 (S96C/S99C/Y102C/D207A/I208R/F209S) are introduced into Saccharomyces cerevisiae, which already contain the Pichia stipitis XYL1 gene (encoding xylose reductase) and the endogenously overexpressed XKS1 gene (encoding xylulokinase) Scheffersomyces stipitis

Protein Variants

Protein Variants Comment Organism
D207A/I208R/F209S mutant bearing a reversal of coenzyme specificity from NAD+ to NADP+ is introduced into Saccharomyces cerevisiae. kcat/Km (NAD+): 181 l/min/mmol, kcat/Km (NADP+): 2790 l/min/mmol, Km (xylitol in the presence of NAD+): 31.1 mM, NADP+-dependent activity: 0.782 U/mg, NAD+-dependent activity: 0.271 U/mg. In xylose fermentation a large decrease in xylitol and glycerol yield is shown, while the xylose consumption and ethanol yield are decreased Scheffersomyces stipitis
S96C/S99C/Y102C/D207A/I208R/F209S mutant bearing a reversal of coenzyme specificity from NAD+ to NADP+ and additional zinc-binding site for thermostability, is introduced into Saccharomyces cerevisiae. kcat/Km (NADP+): 10700 l/min/mmol, Km (xylitol in the presence of NAD+): 111 mM , NADP+-dependent activity: 0.689 U/mg, NAD+-dependent activity: 0.136 U/mg. The xylose consumption and ethanol yield are decreased, and the xylitol yield is increased, because of low XDH activity Scheffersomyces stipitis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
21.7
-
 xylitol wild-type XDH: kcat/Km (NAD+): 2760 l/min/mmol, kcat/Km (NADP+): 2790 l/min/mmol Scheffersomyces stipitis
31.1
-
 xylitol mutant D207A/I208R/F209S: kcat/Km (NAD+): 181 l/min/mmol, kcat/Km (NADP+): 0.65 l/min/mmol Scheffersomyces stipitis
111
-
 xylitol mutant S96C/S99C/Y102C/D207A/I208R/F209S: kcat/Km (NADP+): 10700 l/min/mmol Scheffersomyces stipitis

Organism

Organism UniProt Comment Textmining
Scheffersomyces stipitis
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 wild-type: NADP+-dependent activity: 0.005 U/mg, NAD+-dependent activity: 1.654 U/mg, mutant D207A/I208R/F209S: NADP+-dependent activity: 0.782 U/mg, NAD+-dependent activity: 0.271 U/mg, mutant S96C/S99C/Y102C/D207A/I208R/F209S: NADP+-dependent activity: 0.698 U/mg, NAD+-dependent activity: 0.136 U/mg Scheffersomyces stipitis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
xylitol + NAD+
-
 Scheffersomyces stipitis D-xylulose + NADH + H+
-
 ?
xylitol + NADP+
-
 Scheffersomyces stipitis D-xylulose + NADPH + H+
-
 ?

Synonyms

Synonyms Comment Organism
XDH
-
 Scheffersomyces stipitis
XL2
-
 Scheffersomyces stipitis
xylitol dehydrogenase
-
 Scheffersomyces stipitis