Cloned (Comment) | Organism |
---|---|
derivatives of the expression vector pQE32 containing wild-type and mutated versions of ladA transformed to Escherichia coli M13 cells | Aspergillus niger |
Crystallization (Comment) | Organism |
---|---|
modelling of LadA on human SDH identifies residues M70 and Y318 in LadA, that may explain the absence of activity on D-sorbitol | Aspergillus niger |
Protein Variants | Comment | Organism |
---|---|---|
M70F | almost complete loss of activity | Aspergillus niger |
M70F | results in a nearly complete enzyme inactivation | Aspergillus niger |
Y318F | results in a significant increase in affinity for D-sorbitol, xylitol and L-arabitol | Aspergillus niger |
Y318F | increased activity for L-arabinitol and xylitol, increased affinity for D-sorbitol | Aspergillus niger |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.056 | - |
L-arabitol | wild-type | Aspergillus niger | |
0.056 | - |
L-arabinitol | wild-type, pH 9.6, 20°C | Aspergillus niger | |
0.078 | - |
L-arabitol | mutant Y318F | Aspergillus niger | |
0.078 | - |
L-arabinitol | mutant Y318F, pH 9.6, 20°C | Aspergillus niger | |
0.218 | - |
xylitol | mutant Y318F | Aspergillus niger | |
0.218 | - |
xylitol | mutant Y318F, pH 9.6, 20°C | Aspergillus niger | |
0.25 | - |
xylitol | wild-type | Aspergillus niger | |
0.25 | - |
xylitol | wild-type, pH 9.6, 20°C | Aspergillus niger | |
0.868 | - |
D-sorbitol | mutant Y318F, pH 9.6, 20°C | Aspergillus niger | |
4.12 | - |
D-sorbitol | wild-type, pH 9.6, 20°C | Aspergillus niger | |
4.122 | - |
D-sorbitol | wild-type | Aspergillus niger | |
4.868 | - |
D-sorbitol | mutant Y318F | Aspergillus niger |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aspergillus niger | A2QAC0 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant proteins purified using Ni-agarose | Aspergillus niger |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-sorbitol + NAD+ | - |
Aspergillus niger | D-fructose + NADH + H+ | - |
? | |
D-sorbitol + NAD+ | wild-type shows no activity on D-sorbitol, whereas mutant Y318F does | Aspergillus niger | ? + NADH + H+ | - |
? | |
L-arabinitol + NAD+ | - |
Aspergillus niger | L-xylulose + NADH + H+ | - |
? | |
L-arabitol + NAD+ | - |
Aspergillus niger | L-xylulose + NADH + H+ | - |
? | |
additional information | Y318 of LadA contributes significantly to the substrate specificity difference between LAD and xylitol dehydrogenase/D-sorbitol dehydrogenase | Aspergillus niger | ? | - |
? | |
xylitol + NAD+ | - |
Aspergillus niger | D-xylulose + NADH + H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
L-arabitol dehydrogenase | - |
Aspergillus niger |
LAD | - |
Aspergillus niger |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
13.48 | - |
D-sorbitol | wild-type, pH 9.6, 20°C | Aspergillus niger | |
13.88 | - |
D-sorbitol | mutant Y318F, pH 9.6, 20°C | Aspergillus niger | |
14.4 | - |
L-arabinitol | wild-type, pH 9.6, 20°C | Aspergillus niger | |
19.67 | - |
xylitol | wild-type, pH 9.6, 20°C | Aspergillus niger | |
30 | - |
L-arabinitol | mutant Y318F, pH 9.6, 20°C | Aspergillus niger | |
36.8 | - |
xylitol | mutant Y318F, pH 9.6, 20°C | Aspergillus niger | |
809 | - |
D-sorbitol | wild-type | Aspergillus niger | |
833 | - |
D-sorbitol | mutant Y318F | Aspergillus niger | |
863 | - |
L-arabitol | wild-type | Aspergillus niger | |
1180 | - |
xylitol | wild-type | Aspergillus niger | |
1800 | - |
L-arabitol | mutant Y318F | Aspergillus niger | |
2208 | - |
xylitol | mutant Y318F | Aspergillus niger |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Aspergillus niger |