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 (Comment) | Organism |
---|---|
expression in Escherichia coli | Geobacillus stearothermophilus |
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 | Comment | Organism | Structure |
---|---|---|---|
Butyraldehyde | - |
Geobacillus stearothermophilus |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus stearothermophilus | P42328 | - |
- |
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 | 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 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 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 | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Geobacillus stearothermophilus |
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 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 |