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Literature summary for 1.1.1.383 extracted from

  • Bastian, S.; Liu, X.; Meyerowitz, J.; Snow, C.; Chen, M.; Arnold, F.
    Engineered ketol-acid reductoisomerase and alcohol dehydrogenase enable anaerobic 2-methylpropan-1-ol production at theoretical yield in Escherichia coli (2011), Metab. Eng., 13, 345-352.
    View publication on PubMed

Application

Application Comment Organism
biofuel production Recent work has demonstrated glucose to isobutanol conversion through a modified amino acid pathway in a recombinant organism. We demonstrate that an NADH-dependent pathway enables anaerobic isobutanol production at 100% theoretical yield and at higher titer and productivity than both the NADPH-dependent pathway and transhydrogenase over-expressing strain Escherichia coli

Protein Variants

Protein Variants Comment Organism
A71S mutant engineered to use NADH instead of NADPH Escherichia coli
A71S/R76D/S78D/Q110A mutant engineered to use NADH instead of NADPH Escherichia coli
A71S/R76D/S78D/Q110V mutant engineered to use NADH instead of NADPH Escherichia coli
Q110A mutant engineered to use NADH instead of NADPH Escherichia coli
Q110V mutant engineered to use NADH instead of NADPH Escherichia coli
R76D mutant engineered to use NADH instead of NADPH Escherichia coli
S78D mutant engineered to use NADH instead of NADPH Escherichia coli
S78D/Q110V mutant engineered to use NADH instead of NADPH Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.013
-
NADPH mutant Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
0.024
-
NADPH mutant Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
0.03
-
NADH mutant A71S/R76D/S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
0.04
-
NADPH wild-type, pH 7.0, temperature not specified in the publication Escherichia coli
0.04
-
NADH mutant A71S/R76D/S78D/Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
0.065
-
NADPH mutant S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
0.065
-
NADH mutant S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
0.13
-
NADH mutant S78D, pH 7.0, temperature not specified in the publication Escherichia coli
0.14
-
NADH mutant Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
0.28
-
NADH mutant Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
0.57
-
NADPH mutant A71S/R76D/S78D/Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
0.65
-
NADPH mutant A71S/R76D/S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
0.66
-
NADPH mutant S78D, pH 7.0, temperature not specified in the publication Escherichia coli
1.08
-
NADH wild-type, pH 7.0, temperature not specified in the publication Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P05793
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-2-acetolactate + NADH + H+
-
Escherichia coli 2,3-dihydroxy-3-methylbutanoate + NAD+
-
?
(S)-2-acetolactate + NADPH + H+
-
Escherichia coli 2,3-dihydroxy-3-methylbutanoate + NADP+
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.2
-
NADPH mutant A71S/R76D/S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
0.3
-
NADH wild-type, pH 7.0, temperature not specified in the publication Escherichia coli
0.6
-
NADPH mutant A71S/R76D/S78D/Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
2
-
NADH mutant S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
2.2
-
NADPH mutant S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
2.3
-
NADH mutant A71S/R76D/S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
2.5
-
NADPH mutant S78D, pH 7.0, temperature not specified in the publication Escherichia coli
2.6
-
NADH mutant A71S/R76D/S78D/Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
3.1
-
NADH mutant Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
3.3
-
NADH mutant Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
3.6
-
NADPH wild-type, pH 7.0, temperature not specified in the publication Escherichia coli
3.6
-
NADH mutant S78D, pH 7.0, temperature not specified in the publication Escherichia coli
7.2
-
NADPH mutant Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
7.2
-
NADPH mutant Q110V, pH 7.0, temperature not specified in the publication Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.3
-
NADH wild-type, pH 7.0, temperature not specified in the publication Escherichia coli
0.5
-
NADPH mutant A71S/R76D/S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
1.1
-
NADPH mutant A71S/R76D/S78D/Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
3.8
-
NADPH mutant S78D, pH 7.0, temperature not specified in the publication Escherichia coli
11
-
NADH mutant Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
24.8
-
NADH mutant Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
27.6
-
NADH mutant S78D, pH 7.0, temperature not specified in the publication Escherichia coli
31.8
-
NADH mutant S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
34.2
-
NADPH mutant S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
60.3
-
NADH mutant A71S/R76D/S78D/Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
74
-
NADH mutant A71S/R76D/S78D/Q110V, pH 7.0, temperature not specified in the publication Escherichia coli
87.3
-
NADPH wild-type, pH 7.0, temperature not specified in the publication Escherichia coli
302
-
NADPH mutant Q110A, pH 7.0, temperature not specified in the publication Escherichia coli
569
-
NADPH mutant Q110V, pH 7.0, temperature not specified in the publication Escherichia coli