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Literature summary extracted from

  • Loder, A.J.; Zeldes, B.M.; Garrison, G.D.; Lipscomb, G.L.; Adams, M.W.; Kelly, R.M.
    Alcohol selectivity in a synthetic thermophilic n-butanol pathway is driven by biocatalytic and thermostability characteristics of constituent enzymes (2015), Appl. Environ. Microbiol., 81, 7187-7200.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
1.1.1.1 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.
1.2.1.10 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.
1.2.1.10 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(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.1 expression in Escherichia coli Thermoanaerobacter sp.
1.2.1.10 expression in Escherichia coli Thermoanaerobacter sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.10 0.0057
-
 butyryl-CoA pH 7.9, 60°C Thermoanaerobacter sp.
1.2.1.10 0.0094
-
 acetyl-CoA pH 7.9, 60°C Thermoanaerobacter sp.
1.2.1.10 0.0272
-
 butyryl-CoA pH 7.9, 60°C Thermoanaerobacter sp.
1.2.1.10 0.0836
-
 acetyl-CoA pH 7.9, 60°C Thermoanaerobacter sp.
1.2.1.10 3.23
-
 Butyraldehyde pH 7.9, 60°C Thermoanaerobacter sp.
1.2.1.10 22
-
 acetaldehyde pH 7.9, 60°C Thermoanaerobacter sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.2.1.10 51700
-
 5 * 51700, calculated, 5 * 52400, SDS-PAGE Thermoanaerobacter sp.
1.2.1.10 52400
-
 5 * 51700, calculated, 5 * 52400, SDS-PAGE Thermoanaerobacter sp.
1.2.1.10 94900
-
 6 * 97700, calculated, 6 * 94900, SDS-PAGE Thermoanaerobacter sp.
1.2.1.10 97700
-
 6 * 97700, calculated, 6 * 94900, SDS-PAGE Thermoanaerobacter sp.
1.2.1.10 265000
-
 gel filtration Thermoanaerobacter sp.
1.2.1.10 555000
-
 gel filtration Thermoanaerobacter sp.

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.1 Thermoanaerobacter sp. B0K4A2
-
-
1.2.1.10 Thermoanaerobacter sp. B0K315
-
-
1.2.1.10 Thermoanaerobacter sp. B0K4A2
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.10 acetaldehyde + CoA + NAD+
-
 Thermoanaerobacter sp. acetyl-CoA + NADH + H+
-
 r
1.2.1.10 acetyl-CoA + NADH + H+
-
 Thermoanaerobacter sp. acetaldehyde + CoA + NAD+
-
 r
1.2.1.10 butyraldehyde + CoA + NAD+
-
 Thermoanaerobacter sp. butyryl-CoA + NADH + H+
-
 r
1.2.1.10 butyryl-CoA + NADH + H+
-
 Thermoanaerobacter sp. butyraldehyde + CoA + NAD+
-
 r

Subunits

EC Number Subunits Comment Organism
1.2.1.10 hexamer 6 * 97700, calculated, 6 * 94900, SDS-PAGE Thermoanaerobacter sp.
1.2.1.10 pentamer 5 * 51700, calculated, 5 * 52400, SDS-PAGE Thermoanaerobacter sp.

Synonyms

EC Number Synonyms Comment Organism
1.1.1.1 AdhE bifunctional enzyme, catalyzes reactions of EC 1.1.1.1 and EC 1.2.1.10 Thermoanaerobacter sp.
1.1.1.1 Teth514_0627
-
 Thermoanaerobacter sp.
1.2.1.10 AdhE bifunctional enzyme, catalyzes reactions of EC 1.1.1.1 and EC 1.2.1.10 Thermoanaerobacter sp.
1.2.1.10 BAD
-
 Thermoanaerobacter sp.
1.2.1.10 Teth514_0627
-
 Thermoanaerobacter sp.
1.2.1.10 Teth514_1942
-
 Thermoanaerobacter sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.10 NAD+
-
 Thermoanaerobacter sp.
1.2.1.10 NADH
-
 Thermoanaerobacter sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.2.1.10 1.09
-
 acetaldehyde pH 7.9, 60°C Thermoanaerobacter sp.
1.2.1.10 7.11
-
 Butyraldehyde pH 7.9, 60°C Thermoanaerobacter sp.
1.2.1.10 55.3
-
 acetyl-CoA pH 7.9, 60°C Thermoanaerobacter sp.
1.2.1.10 381
-
 acetyl-CoA pH 7.9, 60°C Thermoanaerobacter sp.
1.2.1.10 394
-
 butyryl-CoA pH 7.9, 60°C Thermoanaerobacter sp.
1.2.1.10 833
-
 butyryl-CoA pH 7.9, 60°C Thermoanaerobacter sp.