Application | Comment | Organism |
---|---|---|
synthesis | recombinant Escherichia coli expressing mutant N126H/T130K or wild-type synthesizes 66.50 mM and 26.09 mM 4-hydroxyisoleucine, respectively, in 24 h | Bacillus mycoides |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Bacillus mycoides |
Protein Variants | Comment | Organism |
---|---|---|
N126H | mutant obtained by directed evolution, 2.8fold increase in activity | Bacillus mycoides |
N126H/T130K | catalytic efficiency is 2.4fold and 3.0fold higher than wild-type with Ile and 2-oxoglutaric acid as substrates | Bacillus mycoides |
T130K | mutant obtained by site-specific mutagenesis, 1.7fold increase in activity | Bacillus mycoides |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.5 | - |
2-oxoglutarate | mutant N126H, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.52 | - |
L-isoleucine | mutant N126H, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.85 | - |
2-oxoglutarate | mutant N126H/T130K, pH 7, temperature not specified in the publication | Bacillus mycoides | |
1.51 | - |
L-isoleucine | mutant N126H/T130K, pH 7, temperature not specified in the publication | Bacillus mycoides | |
2.01 | - |
2-oxoglutarate | mutant T130K, pH 7, temperature not specified in the publication | Bacillus mycoides | |
2.38 | - |
L-isoleucine | wild-type, pH 7, temperature not specified in the publication | Bacillus mycoides | |
2.76 | - |
2-oxoglutarate | wild-type, pH 7, temperature not specified in the publication | Bacillus mycoides | |
3.34 | - |
L-isoleucine | mutant T130K, pH 7, temperature not specified in the publication | Bacillus mycoides |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus mycoides | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-isoleucine + 2-oxoglutarate + O2 | - |
Bacillus mycoides | (4S)-4-hydroxy-L-isoleucine + succinate + CO2 | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.289 | - |
L-isoleucine | mutant N126H, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.309 | - |
2-oxoglutarate | mutant T130K, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.318 | - |
2-oxoglutarate | mutant N126H, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.481 | - |
L-isoleucine | wild-type, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.502 | - |
L-isoleucine | mutant T130K, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.527 | - |
2-oxoglutarate | mutant N126H/T130K, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.577 | - |
2-oxoglutarate | wild-type, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.736 | - |
L-isoleucine | mutant N126H/T130K, pH 7, temperature not specified in the publication | Bacillus mycoides |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.15 | - |
L-isoleucine | mutant T130K, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.154 | - |
2-oxoglutarate | mutant T130K, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.203 | - |
L-isoleucine | wild-type, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.209 | - |
2-oxoglutarate | wild-type, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.449 | - |
L-isoleucine | mutant N126H/T130K, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.56 | - |
L-isoleucine | mutant N126H, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.62 | - |
2-oxoglutarate | mutant N126H/T130K, pH 7, temperature not specified in the publication | Bacillus mycoides | |
0.637 | - |
2-oxoglutarate | mutant N126H, pH 7, temperature not specified in the publication | Bacillus mycoides |