Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli JM 109 | Candida sp. HA 167 |
Protein Variants | Comment | Organism |
---|---|---|
E154C | mutant bearing a disrupted Zn2+ binding site: purified preparations show a variable Zn2+ (0.10-0.40 atom/subunit), mutant exhibits a constant catalytic Zn2+ centre activity and does not require exogenous Zn2+ for activity or stability. E154C retains 0.019% and 0.74% of wild-type catalytic efficiency (kcat/Km (sorbitol): 7800/Msec and kcat:161/sec) for NAD+-dependent oxidation of sorbitol at 25°C respectively. The pH profile of kcat/Ksorbitol for E154C decreases below an apparent pK of 9.1, reflecting a shift in pK by about +1.7-1.9 pH units compared with the corresponding pH profiles for wild-type. IC50 (ZnSO4): 0.005 mM | Candida sp. HA 167 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | exogenous Zn2+ (added as ZnSO4) in the concentration range 1-100 microM is a strong irreversible inhibitor of wild-type and mutant E154C | Candida sp. HA 167 | |
ZnSO4 | - |
Candida sp. HA 167 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3 | - |
D-sorbitol | kcat/Km (sorbitol): 9500/Msec | Ovis aries | |
21 | - |
sorbitol | kcat/Km (sorbitol): 7800/Msec | Candida sp. HA 167 | |
290 | - |
NAD+ | mutant E154C, kcat/Km (NAD+): 4100/Msec | Candida sp. HA 167 | |
430 | - |
NAD+ | kcat/Km (NAD+): 370000/Msec | Candida sp. HA 167 | |
500 | - |
NAD+ | kcat/Km (NAD+): 57000/Msec | Ovis aries | |
785 | - |
sorbitol | mutant E154C, kcat/Km (sorbitol): 1.5/Msec | Candida sp. HA 167 |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
160000 | - |
gel filtration, protein is a homotetramer | Candida sp. HA 167 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Candida sp. HA 167 | - |
- |
- |
Ovis aries | - |
- |
- |
Purification (Comment) | Organism |
---|---|
using affinity chromatography and preparative gel filtration | Candida sp. HA 167 |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
liver | - |
Ovis aries | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-sorbitol + NAD+ | - |
Ovis aries | D-fructose + NADH + H+ | - |
r | |
sorbitol + NAD+ | - |
Candida sp. HA 167 | D-fructose + NADH + H+ | - |
? | |
xylitol + NAD+ | - |
Ovis aries | D-xylulose + NADH + H+ | - |
? | |
xylitol + NAD+ | - |
Candida sp. HA 167 | D-xylulose + NADH + H+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | gel filtration | Candida sp. HA 167 |
Synonyms | Comment | Organism |
---|---|---|
GmXDH | - |
Candida sp. HA 167 |
slSDH | - |
Ovis aries |
xylitol dehydrogenase | - |
Ovis aries |
xylitol dehydrogenase | - |
Candida sp. HA 167 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Ovis aries |
25 | - |
assay at | Candida sp. HA 167 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.19 | - |
sorbitol | mutant E154C | Candida sp. HA 167 | |
2 | 8 | D-sorbitol | - |
Ovis aries | |
161 | - |
sorbitol | - |
Candida sp. HA 167 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
assay at | Ovis aries |
9 | - |
assay at | Candida sp. HA 167 |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.005 | - |
value identical for wildtype and for mutant E154C | Candida sp. HA 167 | ZnSO4 |