Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.1 extracted from

  • Kirmair, L.; Seiler, D.L.; Skerra, A.
    Stability engineering of the Geobacillus stearothermophilus alcohol dehydrogenase and application for the synthesis of a polyamide 12 precursor (2015), Appl. Microbiol. Biotechnol., 99, 10501-10513.
    View publication on PubMed

Application

Application Comment Organism
synthesis semi-preparative biocatalysis at 60°C using the stabilized mutant C257L, employing butyraldehyde for in situ cofactor regeneration with only catalytic amounts of NAD+, yields up to 23% conversion of omega-hydroxy lauric acid methyl ester to omega-oxo lauric acid methyl ester after 30 min Geobacillus stearothermophilus

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Geobacillus stearothermophilus

Protein Variants

Protein Variants Comment Organism
C257L mutation introduced to improve stability under oxidzing conditions. Mutant exhibits prolonged stability and an elevated inactivation temperature Geobacillus stearothermophilus

Inhibitors

Inhibitors Comment Organism Structure
Butyraldehyde
-
Geobacillus stearothermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.066
-
12-oxolauric acid methyl ester wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
0.072
-
NADH wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
0.086
-
12-hydroxylauric acid methyl ester wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
0.1
-
12-hydroxylauric acid methyl ester mutant C257L, pH 8.0, 60°C Geobacillus stearothermophilus
0.108
-
Butyraldehyde wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
0.239
-
NAD+ wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
0.364
-
acetaldehyde wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
0.611
-
n-butanol wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
0.91
-
ethanol wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
1.5
-
ethanol mutant C257L, pH 8.0, 60°C Geobacillus stearothermophilus

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus P42328
-
-

Storage Stability

Storage Stability Organism
15°C, 50 mM sodium phosphate buffer, pH 8.0, half-life of wild-type 7 h, half-life of mutant C257L 17 h Geobacillus stearothermophilus
4°C, 50 mM sodium phosphate buffer, pH 7.0, 10 mM 2-mercaptoethanol, 10% glycerol, both wild-type and mutant C357L stable for several months Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
12-hydroxylauric acid methyl ester + NAD+
-
Geobacillus stearothermophilus 12-oxolauric acid methyl ester + NADH + H+ product is a key intermediate for biobased polyamide 12 production ?
12-oxolauric acid methyl ester + NADH + H+
-
Geobacillus stearothermophilus 12-hydroxylauric acid methyl ester + NAD+
-
?
acetaldehyde + NADH + H+
-
Geobacillus stearothermophilus ethanol + NAD+
-
r
butyraldehyde + NAD+
-
Geobacillus stearothermophilus n-butanol + NADH + H+
-
r
ethanol + NAD+
-
Geobacillus stearothermophilus acetaldehyde + NADH + H+
-
r
n-butanol + NADH + H+
-
Geobacillus stearothermophilus butyraldehyde + NAD+
-
r

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
62.6
-
wild-type, thermal denaturation midpoint Geobacillus stearothermophilus
65.3
-
mutant C257L, thermal denaturation midpoint Geobacillus stearothermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
27.6
-
12-hydroxylauric acid methyl ester wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
34.2
-
NAD+ wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
34.3
-
12-hydroxylauric acid methyl ester mutant C257L, pH 8.0, 60°C Geobacillus stearothermophilus
195
-
n-butanol wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
305
-
ethanol wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
308
-
12-oxolauric acid methyl ester wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
326
-
NADH wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
365
-
ethanol mutant C257L, pH 8.0, 60°C Geobacillus stearothermophilus
681
-
acetaldehyde wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
931
-
Butyraldehyde wild-type, pH 8.0, 60°C Geobacillus stearothermophilus

Cofactor

Cofactor Comment Organism Structure
NADH
-
Geobacillus stearothermophilus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.1
-
Butyraldehyde wild-type, pH 8.0, 60°C Geobacillus stearothermophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
100
-
NAD+ wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
240
-
ethanol mutant C257L, pH 8.0, 60°C Geobacillus stearothermophilus
320
-
n-butanol wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
320
-
12-hydroxylauric acid methyl ester wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
340
-
12-hydroxylauric acid methyl ester mutant C257L, pH 8.0, 60°C Geobacillus stearothermophilus
340
-
ethanol wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
1900
-
acetaldehyde wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
4500
-
NADH wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
4700
-
12-oxolauric acid methyl ester wild-type, pH 8.0, 60°C Geobacillus stearothermophilus
8600
-
Butyraldehyde wild-type, pH 8.0, 60°C Geobacillus stearothermophilus