Application | Comment | Organism |
---|---|---|
synthesis | synthetic pathway for n-butanol production from acetyl coenzyme at 70°C, using beta-ketothiolase Thl, 3-hydroxybutyryl-CoA dehydrogenase Hbd, and 3-hydroxybutyryl-CoA dehydratase Crt from Caldanaerobacter subterraneus subsp. tengcongensis, trans-2-enoyl-CoA reductase Ter from Spirochaeta thermophila and bifunctional aldehyde dehydrogenase AdhE and and butanol dehydrogenase in vitro. n-Butanol is produced at 70°C, but with different amounts of ethanol as a coproduct, because of the broad substrate specificities of AdhE, Bad, and Bdh. A reaction kinetics model, validated via comparison to in vitro experiments, is used to determine relative enzyme ratios needed to maximize n-butanol production. By using large relative amounts of Thl and Hbd and small amounts of Bad and Bdh, >70% conversion to n-butanol is observed in vitro, but with a 60% decrease in the predicted pathway flux | Thermoanaerobacter sp. |
synthesis | synthetic pathway for n-butanol production from acetyl-CoA at 70°C, using beta-ketothiolase Thl, 3-hydroxybutyryl-CoA dehydrogenase Hbd, and 3-hydroxybutyryl-CoA dehydratase Crt from Caldanaerobacter subterraneus subsp. tengcongensis, trans-2-enoyl-CoA reductase Ter from Spirochaeta thermophila and bifunctional aldehyde dehydrogenase AdhE and and butanol dehydrogenase in vitro. n-Butanol is produced at 70°C, but with different amounts of ethanol as a coproduct, because of the broad substrate specificities of AdhE, Bad, and Bdh. A reaction kinetics model, validated via comparison to in vitro experiments, is used to determine relative enzyme ratios needed to maximize n-butanol production. By using large relative amounts of Thl and Hbd and small amounts of Bad and Bdh, >70% conversion to n-butanol is observed in vitro, but with a 60% decrease in the predicted pathway flux | Thermoanaerobacter sp. |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Thermoanaerobacter sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0057 | - |
butyryl-CoA | pH 7.9, 60°C | Thermoanaerobacter sp. | |
0.0094 | - |
acetyl-CoA | pH 7.9, 60°C | Thermoanaerobacter sp. | |
0.0272 | - |
butyryl-CoA | pH 7.9, 60°C | Thermoanaerobacter sp. | |
0.0836 | - |
acetyl-CoA | pH 7.9, 60°C | Thermoanaerobacter sp. | |
3.23 | - |
Butyraldehyde | pH 7.9, 60°C | Thermoanaerobacter sp. | |
22 | - |
acetaldehyde | pH 7.9, 60°C | Thermoanaerobacter sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
51700 | - |
5 * 51700, calculated, 5 * 52400, SDS-PAGE | Thermoanaerobacter sp. |
52400 | - |
5 * 51700, calculated, 5 * 52400, SDS-PAGE | Thermoanaerobacter sp. |
94900 | - |
6 * 97700, calculated, 6 * 94900, SDS-PAGE | Thermoanaerobacter sp. |
97700 | - |
6 * 97700, calculated, 6 * 94900, SDS-PAGE | Thermoanaerobacter sp. |
265000 | - |
gel filtration | Thermoanaerobacter sp. |
555000 | - |
gel filtration | Thermoanaerobacter sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermoanaerobacter sp. | B0K315 | - |
- |
Thermoanaerobacter sp. | B0K4A2 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetaldehyde + CoA + NAD+ | - |
Thermoanaerobacter sp. | acetyl-CoA + NADH + H+ | - |
r | |
acetyl-CoA + NADH + H+ | - |
Thermoanaerobacter sp. | acetaldehyde + CoA + NAD+ | - |
r | |
butyraldehyde + CoA + NAD+ | - |
Thermoanaerobacter sp. | butyryl-CoA + NADH + H+ | - |
r | |
butyryl-CoA + NADH + H+ | - |
Thermoanaerobacter sp. | butyraldehyde + CoA + NAD+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
hexamer | 6 * 97700, calculated, 6 * 94900, SDS-PAGE | Thermoanaerobacter sp. |
pentamer | 5 * 51700, calculated, 5 * 52400, SDS-PAGE | Thermoanaerobacter sp. |
Synonyms | Comment | Organism |
---|---|---|
AdhE | bifunctional enzyme, catalyzes reactions of EC 1.1.1.1 and EC 1.2.1.10 | Thermoanaerobacter sp. |
BAD | - |
Thermoanaerobacter sp. |
Teth514_0627 | - |
Thermoanaerobacter sp. |
Teth514_1942 | - |
Thermoanaerobacter sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Thermoanaerobacter sp. | |
NADH | - |
Thermoanaerobacter sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.09 | - |
acetaldehyde | pH 7.9, 60°C | Thermoanaerobacter sp. | |
7.11 | - |
Butyraldehyde | pH 7.9, 60°C | Thermoanaerobacter sp. | |
55.3 | - |
acetyl-CoA | pH 7.9, 60°C | Thermoanaerobacter sp. | |
381 | - |
acetyl-CoA | pH 7.9, 60°C | Thermoanaerobacter sp. | |
394 | - |
butyryl-CoA | pH 7.9, 60°C | Thermoanaerobacter sp. | |
833 | - |
butyryl-CoA | pH 7.9, 60°C | Thermoanaerobacter sp. |