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Literature summary for 1.1.1.12 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 (2017), J. Biosci. Bioeng., 123, 20-27 .
    View publication on PubMed
Search for more literature for 1.1.1.12

Cloned(Commentary)

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

Inhibitors

Inhibitors Comment Organism Structure
diphenylhydantoine moderate inhibition Meyerozyma caribbica
DTT slight inhibition Meyerozyma caribbica
EDTA strong inhibition Meyerozyma caribbica
EGTA moderate inhibition Meyerozyma caribbica
furfural moderate inhibition Meyerozyma caribbica
iodoacetamide moderate inhibition Meyerozyma caribbica
iodoacetic acid strong inhibition Meyerozyma caribbica
o-phenanthroline strong inhibition Meyerozyma caribbica
potassium ferricyanide K4[Fe(CN)6], moderate inhibition Meyerozyma caribbica
Sodium fluoride moderate inhibition Meyerozyma caribbica
Tiron strong inhibition Meyerozyma caribbica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1
-
 NAD+ pH 7.0, 30°C, recombinant enzyme, with xylitol Meyerozyma caribbica
0.11
-
 NAD+ pH 7.0, 30°C, native enzyme, with xylitol Meyerozyma caribbica
7.28
-
 NADP+ pH 7.0, 30°C, native enzyme, with xylitol Meyerozyma caribbica
15.8
-
 ribitol pH 7.0, 30°C, native enzyme Meyerozyma caribbica
16
-
 xylitol pH 7.0, 30°C, recombinant enzyme Meyerozyma caribbica
16.1
-
 xylitol pH 7.0, 30°C, native enzyme Meyerozyma caribbica
21.2
-
 D-sorbitol pH 7.0, 30°C, native enzyme Meyerozyma caribbica
31.1
-
 L-arabitol pH 7.0, 30°C, native enzyme Meyerozyma caribbica
132
-
 L-arabitol pH 7.0, 30°C, recombinant enzyme Meyerozyma caribbica

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ activates Meyerozyma caribbica
Fe2+ activates Meyerozyma caribbica
additional information after incubation with ZnSO4, ZnCl2, FeCl2, and CuCl2, 40, 37, 30, and 25%, respectively, of the activity of the metal-free enzyme is restored. Poorer effects by Fe3+, Cd2+, and Ca2+, while Ni2+, Mn2+, Co2+, Mg2+, K+, and Na+ do not exert restorative effects Meyerozyma caribbica
Zn2+ required, the zinc ADH signature and conserved coenzyme binding motif are observed in the amino acid sequence of McXDH at positions 65-76 and 183-188 and are completely conserved among McXDH and XDHs from other yeasts and filamentous fungi Meyerozyma caribbica

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
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+
-
 r

Organism

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

Purification (Commentary)

Purification (Comment) Organism
native enzyme by dialysis, anion exchange chromatography, ammoniuim sulfate fractionation, hydrophobic interaction chromatography, again dialysis and another different step off anion exchange chromatography, dialysis, and gel filtration, followed by repeating the last anion exchange chromatographic step. Recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography Meyerozyma caribbica

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.4
-
 about, purified recombinant enzyme, substrates L-arabitol and NAD+, pH 7.0, 30°C Meyerozyma caribbica
2
-
 purified recombinant enzyme, substrates xylitol and NAD+, pH 7.0, 30°C Meyerozyma caribbica
4.9
-
 purified native enzyme, substrates L-arabitol and NAD+, pH 7.0, 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
L-arabitol + NAD+ about 20% activity compared to xylitol Meyerozyma caribbica L-xylulose + NADH + H+
-
 r
additional information the enzyme has L-arabitol dehydrogenase (LAD) activity and also exhibits broad specificity to polyols, such as xylitol, D-sorbitol, and ribitol. Xylitol is the preferred substrate (EC 1.1.1.9) Meyerozyma caribbica ?
-
-
ribitol + NAD+
-
 Meyerozyma caribbica D-ribulose + NADH + H+
-
 r
xylitol + NAD+
-
 Meyerozyma caribbica D-xylulose + NADH + H+
-
 r
xylitol + NAD+ preferred substrates, reaction of EC 1.1.1.9 Meyerozyma caribbica D-xylulose + NADH + H+
-
 r
xylitol + NADP+ very low activity with NADP+ Meyerozyma caribbica D-xylulose + NADPH + H+
-
 r

Subunits

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

Synonyms

Synonyms Comment Organism
L-arabitol dehydrogenase
-
 Meyerozyma caribbica
LAD
-
 Meyerozyma caribbica
More see also EC 1.1.1.9 Meyerozyma caribbica
XYL2
-
 Meyerozyma caribbica

Temperature Optimum [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45 60 the enzyme partially loses its activity at 45-55°C and completely loses activity at temperatures greater than 60°C. McXDH exhibits activities of approximately 81% and 51% at 30°C and 35°C after 30 min incubation Meyerozyma caribbica

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.27
-
 NADP+ pH 7.0, 30°C, native enzyme, with xylitol Meyerozyma caribbica
3.35
-
 L-arabitol pH 7.0, 30°C, native enzyme Meyerozyma caribbica
4.48
-
 ribitol pH 7.0, 30°C, native enzyme Meyerozyma caribbica
8.28
-
 D-sorbitol pH 7.0, 30°C, native enzyme Meyerozyma caribbica
18
-
 xylitol pH 7.0, 30°C, native enzyme Meyerozyma caribbica
18.5
-
 NAD+ pH 7.0, 30°C, native enzyme, with xylitol Meyerozyma caribbica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9.5
-
-
 Meyerozyma caribbica

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Meyerozyma caribbica
NADH
-
 Meyerozyma caribbica
NADP+ poor activity Meyerozyma caribbica

General Information

General Information Comment Organism
evolution the enzyme belongs to the medium-chain dehydrogenase/reductase (MDR) superfamily and polyol dehydrogenase (PDH) subfamily. The enzyme contains the typical NAD+-binding motif GxGxxG of MDR family enzymes Meyerozyma caribbica
physiological function the organism catabolizes L-arabinose as well as D-glucose and D-xylose. The highest production amounts of ethanol from D-glucose, xylitol from D-xylose, and L-arabitol from L-arabinose were 0.45 g/g D-glucose, 0.60 g/g D-xylose, and 0.61 g/g L-arabinose with 21.7 g/l ethanol, 20.2 g/l xylitol, and 30.3 g/l L-arabitol, respectively. The enzyme has L-arabitol dehydrogenase (LAD) activity and also exhibits broad specificity to polyols, such as xylitol, D-sorbitol, ribitol, and L-arabitol. Xylitol is the preferred substrate 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 pH 7.0, 30°C, native enzyme Meyerozyma caribbica
0.28
-
 ribitol pH 7.0, 30°C, native enzyme Meyerozyma caribbica
0.39
-
 D-sorbitol pH 7.0, 30°C, native enzyme Meyerozyma caribbica
0.45
-
 NADP+ pH 7.0, 30°C, native enzyme, with xylitol Meyerozyma caribbica
1.12
-
 xylitol pH 7.0, 30°C, native enzyme Meyerozyma caribbica
168.18
-
 NAD+ pH 7.0, 30°C, native enzyme, with xylitol Meyerozyma caribbica