Cloned (Comment) | Organism |
---|---|
site directed mutagenesis | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
- |
Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
K163I | 5.25% activity of wild-type activity | Escherichia coli |
K163R | 63.7% activity of wild-type activity | Escherichia coli |
S146A | 20.3% activity of wild-type activity | Escherichia coli |
S146H | 35.6% activity of wild-type activity | Escherichia coli |
Y159F | no activity | Escherichia coli |
Y159H | 13.3% activity of wild-type activity | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.159 | - |
NAD+ | mutant K163R | Escherichia coli | |
0.223 | - |
cholic acid | mutant K163I | Escherichia coli | |
0.279 | - |
NAD+ | - |
Escherichia coli | |
0.283 | - |
cholic acid | mutant S146H | Escherichia coli | |
0.302 | - |
NAD+ | mutant S146A | Escherichia coli | |
0.329 | - |
NAD+ | mutant K163I | Escherichia coli | |
0.361 | - |
cholic acid | - |
Escherichia coli | |
0.364 | - |
NAD+ | mutant S146H | Escherichia coli | |
0.386 | - |
NAD+ | mutant Y159H | Escherichia coli | |
0.407 | - |
cholic acid | mutant Y159H | Escherichia coli | |
0.415 | - |
cholic acid | mutant S146A | Escherichia coli | |
0.612 | - |
cholic acid | mutant K163R | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
28000 | - |
4 * 28000, SDS-PAGE | Escherichia coli |
107100 | - |
sequence determination | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
cholate + NAD+ = 3alpha,12alpha-dihydroxy-7-oxo-5beta-cholan-24-oate + NADH + H+ | Tyr159 acts as basic catalyst, Lys163 binds to NAD(H) and lowers the pKa value of Tyr159, Ser146 stabilizes the substrate, reaction intermediate and product in catalysis | Escherichia coli |
Subunits | Comment | Organism |
---|---|---|
tetramer | 4 * 28000, SDS-PAGE | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
7.92 | - |
cholic acid | mutant K163I | Escherichia coli | |
20.1 | - |
cholic acid | mutant Y159H | Escherichia coli | |
30.7 | - |
cholic acid | mutant S146A | Escherichia coli | |
53.7 | - |
cholic acid | mutant S146H | Escherichia coli | |
96.2 | - |
cholic acid | mutant K163R | Escherichia coli | |
151 | - |
cholic acid | - |
Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
10 | - |
oxidation of cholic acid | Escherichia coli |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 10 | - |
Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | wild type and mutants | Escherichia coli |