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Literature summary for 1.1.1.9 extracted from

  • Sukpipat, W.; Komeda, H.; Prasertsan, P.; Asano, Y.
    Purification and characterization of xylitol dehydrogenase with L-arabitol dehydrogenase activity from the newly isolated pentose-fermenting yeast Meyerozyma caribbica 5XY2 (2016), J. Biosci. Bioeng., 123, 20-27.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene xyl2, DNA and amino acid sequence determination and analysis, cloning in Escherichia coli strain JM109, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Meyerozyma caribbica

Inhibitors

Inhibitors Comment Organism Structure
EDTA
-
Meyerozyma caribbica
iodoacetic acid
-
Meyerozyma caribbica
o-phenanthroline
-
Meyerozyma caribbica
Tiron
-
Meyerozyma caribbica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1
-
xylitol recombinant enzyme, pH 7.0, 30°C Meyerozyma caribbica
0.11
-
NAD+ at pH 7.0 and 30°C Meyerozyma caribbica
0.11
-
xylitol native enzyme, pH 7.0, 30°C Meyerozyma caribbica
15.4
-
xylitol recombinant enzyme, pH 7.0, 30°C Meyerozyma caribbica
15.8
-
ribitol at pH 7.0 and 30°C Meyerozyma caribbica
16
-
xylitol native enzyme, pH 7.0, 30°C Meyerozyma caribbica
16.1
-
xylitol at pH 7.0 and 30°C Meyerozyma caribbica
21.2
-
D-sorbitol at pH 7.0 and 30°C Meyerozyma caribbica
31.1
-
L-arabitol at pH 7.0 and 30°C Meyerozyma caribbica

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ zinc metalloenzyme Meyerozyma caribbica

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
39000
-
-
Meyerozyma caribbica
195000
-
gel filtration Meyerozyma caribbica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
xylitol + NAD+ Meyerozyma caribbica
-
D-xylulose + NADH + H+
-
?
xylitol + NAD+ Meyerozyma caribbica preferred substrate D-xylulose + NADH + H+
-
?
xylitol + NAD+ Meyerozyma caribbica 5XY2 preferred substrate D-xylulose + NADH + H+
-
?
xylitol + NAD+ Meyerozyma caribbica 5XY2
-
D-xylulose + NADH + H+
-
?

Organism

Organism UniProt Comment Textmining
Meyerozyma caribbica
-
-
-
Meyerozyma caribbica A0A1B4XTS0 pentose-fermenting strain isolated from an alcohol fermentation starter in Thailand
-
Meyerozyma caribbica 5XY2
-
-
-
Meyerozyma caribbica 5XY2 A0A1B4XTS0 pentose-fermenting strain isolated from an alcohol fermentation starter in Thailand
-

Purification (Commentary)

Purification (Comment) Organism
DEAEToyopearl 650 M column chromatography, butyl-Toyopearl 650 M column chromatography, MonoQ column chromatography, and Superdex 200 gel filtration Meyerozyma caribbica
native enzyme by ammonium sulfate fractionation, anion exchange chromatography, hydrophobic interation chromatography, and another different step of anion exchange chromatography, followed by gel filtration. Recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography Meyerozyma caribbica

Source Tissue

Source Tissue Comment Organism Textmining
cell culture strain 5XY2 catabolize L-arabinose as well as D-glucose and D-xylose in fermentation Meyerozyma caribbica
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.08
-
crude enzyme, at pH 7.0 and 30°C Meyerozyma caribbica
2
-
purified recombinant enzyme, pH 7.0, 40°C Meyerozyma caribbica
4.9
-
after purification, at pH 7.0 and 30°C Meyerozyma caribbica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-sorbitol + NAD+
-
Meyerozyma caribbica D-fructose + NADH + H+
-
r
D-sorbitol + NAD+
-
Meyerozyma caribbica 5XY2 D-fructose + NADH + H+
-
r
D-sorbitol + NAD+
-
Meyerozyma caribbica ? + NADH + H+
-
?
D-sorbitol + NAD+
-
Meyerozyma caribbica 5XY2 ? + NADH + H+
-
?
L-arabitol + NAD+ the enzyme also has L-arabitol dehydrogenase activity (20.1% activity compared to xylitol) Meyerozyma caribbica L-xylulose + NADH + H+
-
?
L-arabitol + NAD+ the enzyme also has L-arabitol dehydrogenase activity (20.1% activity compared to xylitol) Meyerozyma caribbica 5XY2 L-xylulose + NADH + H+
-
?
additional information no activity with D-mannitol, erythritol, and D-arabitol. Very low activity with NADP+ Meyerozyma caribbica ?
-
?
additional information the enzyme exhibits broad specificity to polyols, such as xylitol, D-sorbitol, ribitol, and L-arabinitol. Xylitol is the preferred substrate, but native and recombinant enzyme McXDH exhibits relative activities toward L-arabinitol of approximately 20% that toward xylitol. No activity with D-mannitol, erythritol, and D-arabinitol Meyerozyma caribbica ?
-
?
additional information no activity with D-mannitol, erythritol, and D-arabitol. Very low activity with NADP+ Meyerozyma caribbica 5XY2 ?
-
?
additional information the enzyme exhibits broad specificity to polyols, such as xylitol, D-sorbitol, ribitol, and L-arabinitol. Xylitol is the preferred substrate, but native and recombinant enzyme McXDH exhibits relative activities toward L-arabinitol of approximately 20% that toward xylitol. No activity with D-mannitol, erythritol, and D-arabinitol Meyerozyma caribbica 5XY2 ?
-
?
ribitol + NAD+
-
Meyerozyma caribbica ?
-
?
ribitol + NAD+
-
Meyerozyma caribbica 5XY2 ?
-
?
ribitol + NAD+ reaction of EC 1.1.1.56, ribitol 2-dehydrogenase Meyerozyma caribbica D-ribulose + NADH + H+
-
?
ribitol + NAD+ reaction of EC 1.1.1.56, ribitol 2-dehydrogenase Meyerozyma caribbica 5XY2 D-ribulose + NADH + H+
-
?
xylitol + NAD+
-
Meyerozyma caribbica D-xylulose + NADH + H+
-
?
xylitol + NAD+ preferred substrate Meyerozyma caribbica D-xylulose + NADH + H+
-
?
xylitol + NAD+ preferred substrate Meyerozyma caribbica 5XY2 D-xylulose + NADH + H+
-
?
xylitol + NAD+
-
Meyerozyma caribbica 5XY2 D-xylulose + NADH + H+
-
?

Subunits

Subunits Comment Organism
homopentamer
-
Meyerozyma caribbica
homopentamer 5 * 39000, SDS-PAGE Meyerozyma caribbica

Synonyms

Synonyms Comment Organism
XDH
-
Meyerozyma caribbica
XYL2
-
Meyerozyma caribbica
xylitol dehydrogenase
-
Meyerozyma caribbica

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
Meyerozyma caribbica

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
10 55 activity range, inactive at 60°C Meyerozyma caribbica
45 55 the enzyme partially loses its activity at 45-55°C and also completely loses activity at temperatures greater than 60°C Meyerozyma caribbica

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30 45 the enzyme exhibits activities of approximately 81% and 51% after being incubated at 30°C and 35°C, respectively, for 30 min. The enzyme completely loses its activity at temperatures greater than 45°C Meyerozyma caribbica

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.12
-
xylitol recombinant enzyme, pH 7.0, 30°C Meyerozyma caribbica
3.35
-
L-arabitol at pH 7.0 and 30°C Meyerozyma caribbica
4.5
-
ribitol at pH 7.0 and 30°C Meyerozyma caribbica
8.28
-
D-sorbitol at pH 7.0 and 30°C Meyerozyma caribbica
18
-
xylitol at pH 7.0 and 30°C Meyerozyma caribbica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
recombinant enzyme Meyerozyma caribbica
9.5
-
-
Meyerozyma caribbica
9.5
-
native enzyme Meyerozyma caribbica

pH Range

pH Minimum pH Maximum Comment Organism
4 11 the enzyme lost its activity at pH lower than 4.0 and higher than 11.0, corresponding to NAD+ which is unstable at pH values greater than 10.5 Meyerozyma caribbica
4 11 activity range, inactive above or below Meyerozyma caribbica

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Meyerozyma caribbica
NAD+ preferred cofactor Meyerozyma caribbica
NADP+ low activity Meyerozyma caribbica

General Information

General Information Comment Organism
evolution the enzyme belongs to a medium-chain dehydrogenase/reductase (MDR) superfamily and a subfamily of polyol dehydrogenase, PDH Meyerozyma caribbica

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.11
-
L-arabitol at pH 7.0 and 30°C Meyerozyma caribbica
0.29
-
ribitol at pH 7.0 and 30°C Meyerozyma caribbica
0.39
-
D-sorbitol at pH 7.0 and 30°C Meyerozyma caribbica
1.12
-
xylitol at pH 7.0 and 30°C Meyerozyma caribbica