Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Rhizomucor pusillus |
Protein Variants | Comment | Organism |
---|---|---|
D205A/I206R | the mutant shows cofactor preference for NADP+ | Rhizomucor pusillus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ag+ | 9.2% residual activity at 1 mM | Rhizomucor pusillus | |
Ca2+ | complete inhibition at 1 mM | Rhizomucor pusillus | |
Cd2+ | complete inhibition at 1 mM | Rhizomucor pusillus | |
Co2+ | 6.8% residual activity at 1 mM | Rhizomucor pusillus | |
Cu2+ | complete inhibition at 1 mM | Rhizomucor pusillus | |
Fe3+ | 18% residual activity at 1 mM | Rhizomucor pusillus | |
Mg2+ | complete inhibition at 1 mM | Rhizomucor pusillus | |
MgSO4 | complete inhibition at 1 mM | Rhizomucor pusillus | |
Mn2+ | 8.4% residual activity at 1 mM | Rhizomucor pusillus | |
Na+ | complete inhibition at 1 mM | Rhizomucor pusillus | |
Ni2+ | 11.3% residual activity at 1 mM | Rhizomucor pusillus | |
Pb2+ | complete inhibition at 1 mM | Rhizomucor pusillus | |
Zn2+ | 44% residual activity at 1 mM | Rhizomucor pusillus | |
ZnSO4 | 51.2% residual activity at 1 mM | Rhizomucor pusillus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.027 | - |
NAD+ | native enzyme, at pH 7.0 and 30°C | Rhizomucor pusillus | |
0.186 | - |
NAD+ | recombinant enzyme, at pH 7.0 and 30°C | Rhizomucor pusillus | |
0.712 | - |
NADP+ | mutant enzyme D205A/I206R, at pH 7.0 and 30°C | Rhizomucor pusillus | |
2.54 | - |
NADP+ | mutant enzyme D205A, at pH 7.0 and 30°C | Rhizomucor pusillus | |
7.83 | - |
xylitol | native enzyme, at pH 7.0 and 30°C | Rhizomucor pusillus | |
8.25 | - |
D-xylulose | at pH 7.0 and 30°C | Rhizomucor pusillus | |
10.1 | - |
xylitol | recombinant enzyme, at pH 7.0 and 30°C | Rhizomucor pusillus | |
10.2 | - |
NADP+ | recombinant enzyme, at pH 7.0 and 30°C | Rhizomucor pusillus | |
13.5 | - |
NAD+ | mutant enzyme D205A, at pH 7.0 and 30°C | Rhizomucor pusillus | |
19.8 | - |
D-fructose | at pH 7.0 and 30°C | Rhizomucor pusillus | |
31.5 | - |
D-sorbitol | at pH 7.0 and 30°C | Rhizomucor pusillus | |
34.3 | - |
NAD+ | mutant enzyme D205A/I206R, at pH 7.0 and 30°C | Rhizomucor pusillus | |
66.7 | - |
xylitol | mutant enzyme D205A, with NAD+ as cosubstrate, at pH 7.0 and 30°C | Rhizomucor pusillus | |
130 | - |
xylitol | mutant enzyme D205A/I206R, with NADP+ as cosubstrate, at pH 7.0 and 30°C | Rhizomucor pusillus | |
149 | - |
ribitol | at pH 7.0 and 30°C | Rhizomucor pusillus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
41000 | - |
2 * 41000, SDS-PAGE | Rhizomucor pusillus |
87000 | - |
gel filtration | Rhizomucor pusillus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-xylulose + NADH + H+ | Rhizomucor pusillus | - |
xylitol + NAD+ | - |
r | |
D-xylulose + NADH + H+ | Rhizomucor pusillus NBRC 4578 | - |
xylitol + NAD+ | - |
r | |
xylitol + NAD+ | Rhizomucor pusillus | 100% activity | D-xylulose + NADH + H+ | - |
r | |
xylitol + NAD+ | Rhizomucor pusillus NBRC 4578 | 100% activity | D-xylulose + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhizomucor pusillus | S6BFC0 | - |
- |
Rhizomucor pusillus NBRC 4578 | S6BFC0 | - |
- |
Purification (Comment) | Organism |
---|---|
ammonium sulfate fractionation, DEAE Toyopearl chromatography, phenyl Toyopearl chromatography, Mono Q chromatography, Superdex 200 gel filtration, and Reactive Red column chromatography | Rhizomucor pusillus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-fructose + NADH + H+ | - |
Rhizomucor pusillus | D-sorbitol + NAD+ | - |
r | |
D-mannitol + NAD+ | 2.6% activity compared to xylitol | Rhizomucor pusillus | ? | - |
? | |
D-mannitol + NAD+ | 2.6% activity compared to xylitol | Rhizomucor pusillus NBRC 4578 | ? | - |
? | |
D-sorbitol + NAD+ | 71.8% activity compared to xylitol | Rhizomucor pusillus | D-fructose + NADH + H+ | - |
r | |
D-sorbitol + NAD+ | 71.8% activity compared to xylitol | Rhizomucor pusillus NBRC 4578 | D-fructose + NADH + H+ | - |
r | |
D-xylulose + NADH + H+ | - |
Rhizomucor pusillus | xylitol + NAD+ | - |
r | |
D-xylulose + NADH + H+ | - |
Rhizomucor pusillus NBRC 4578 | xylitol + NAD+ | - |
r | |
additional information | the enzyme shows less than 1% activity with erythritol, galactitol, D-arabitol, L-arabitol, and glycerol, and 3.1% activity with NADP+ compared to NAD+ | Rhizomucor pusillus | ? | - |
? | |
additional information | the enzyme shows less than 1% activity with erythritol, galactitol, D-arabitol, L-arabitol, and glycerol, and 3.1% activity with NADP+ compared to NAD+ | Rhizomucor pusillus NBRC 4578 | ? | - |
? | |
ribitol + NAD+ | 60.1% activity compared to xylitol | Rhizomucor pusillus | ? | - |
? | |
xylitol + NAD+ | 100% activity | Rhizomucor pusillus | D-xylulose + NADH + H+ | - |
r | |
xylitol + NAD+ | 100% activity | Rhizomucor pusillus NBRC 4578 | D-xylulose + NADH + H+ | - |
r | |
xylitol + NADP+ | mutant enzyme D205A/I260R shows activity with NADP+ | Rhizomucor pusillus | D-xylulose + NADPH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 41000, SDS-PAGE | Rhizomucor pusillus |
homodimer | 2 * 39185, calculated from amino acid sequence | Rhizomucor pusillus |
Synonyms | Comment | Organism |
---|---|---|
XDH | - |
Rhizomucor pusillus |
XYL2 | - |
Rhizomucor pusillus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
- |
Rhizomucor pusillus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | 50 | approximately 92% and 70% activities remain following incubations at 45 and 50°C, respectively. The enzyme completely loses its activity above 60°C | Rhizomucor pusillus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.185 | - |
NADP+ | recombinant enzyme, at pH 7.0 and 30°C | Rhizomucor pusillus | |
0.33 | - |
NADP+ | mutant enzyme D205A, at pH 7.0 and 30°C | Rhizomucor pusillus | |
0.4 | - |
NADP+ | mutant enzyme D205A/I206R, at pH 7.0 and 30°C | Rhizomucor pusillus | |
1.16 | - |
NAD+ | mutant enzyme D205A/I206R, at pH 7.0 and 30°C | Rhizomucor pusillus | |
5.18 | - |
NAD+ | mutant enzyme D205A, at pH 7.0 and 30°C | Rhizomucor pusillus | |
6.35 | - |
NAD+ | recombinant enzyme, at pH 7.0 and 30°C | Rhizomucor pusillus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Rhizomucor pusillus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 11 | the enzyme loses its activity at pH higher than 11.0 and lower than 4.0 | Rhizomucor pusillus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | dependent on | Rhizomucor pusillus |
Organism | Comment | Expression |
---|---|---|
Rhizomucor pusillus | the enzyme expression levels are increased with accumulation of xylitol | up |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.018 | - |
NADP+ | recombinant enzyme, at pH 7.0 and 30°C | Rhizomucor pusillus | |
0.03 | - |
NAD+ | mutant enzyme D205A/I206R, at pH 7.0 and 30°C | Rhizomucor pusillus | |
0.13 | - |
NADP+ | mutant enzyme D205A, at pH 7.0 and 30°C | Rhizomucor pusillus | |
0.38 | - |
NAD+ | mutant enzyme D205A, at pH 7.0 and 30°C | Rhizomucor pusillus | |
0.56 | - |
NADP+ | mutant enzyme D205A/I206R, at pH 7.0 and 30°C | Rhizomucor pusillus | |
32.36 | - |
NAD+ | recombinant enzyme, at pH 7.0 and 30°C | Rhizomucor pusillus |