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Literature summary for 1.14.11.45 extracted from

  • Huang, S.; Shi, F.
    Directed evolution and site-specific mutagenesis of L-isoleucine dioxygenase derived from Bacillus weihenstephanensis (2018), Biotechnol. Lett., 40, 1227-1235 .
    View publication on PubMed
Search for more literature for 1.14.11.45

Application

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(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Bacillus mycoides

Protein Variants

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 [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

Organism UniProt Comment Textmining
Bacillus mycoides
-
-
-

Substrates and Products (Substrate)

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 [1/s]

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 [mM/s]

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