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Literature summary extracted from

  • Nguyen, D.; Lipscomb, G.; Schut, G.; Vaccaro, B.; Basen, M.; Kelly, R.; Adams, M.
    Temperature-dependent acetoin production by Pyrococcus furiosus is catalyzed by a biosynthetic acetolactate synthase and its deletion improves ethanol production (2016), Metab. Eng., 34, 71-79.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.2.1.6 synthesis the enzyme in Pyrococcus furiosus is a potential platform for the biological production of acetoin at temperatures in the 70-80°C range. Acetoin, or 3-hydroxybutanone, is an important four-carbon compound that serves as a building block for valuable bio-based chemical compounds and is a common flavor additive and preservative in the food industry Pyrococcus furiosus
2.2.1.6 synthesis acetoin (i.e. 3-hydroxybutanone), is a major product at temperatures below 80 °C. Acetolactate synthase ALS, which is involved in branched-chain amino acid biosynthesis, is responsible and deletion of the Als gene abolishes acetoin production. Deletion of Als in a strain of Pyrococcus furiosus heterologously expressing an alcohol dehydrogenase gene from Thermoanaerobacter sp. X514 for ethanol production significantly improves the yield of ethanol Pyrococcus furiosus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.2.1.6 gene als or PF0935, DNA and amino acid sequence determination and analysis Pyrococcus furiosus

Protein Variants

EC Number Protein Variants Comment Organism
2.2.1.6 additional information engineering of a strain of Pyrococcus furiosus to contain an additional pathway for ethanol production. Enzyme ALS deletion abolishes acetoin formation and improves ethanol production in the alcohol dehydrogenase ADHA strain. High level expression of enzyme ALS uncouples the temperature-dependence of acetoin formation Pyrococcus furiosus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.2.1.6 cytoplasm
-
 Pyrococcus furiosus 5737
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.2.1.6 Mg2+ required Pyrococcus furiosus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.2.1.6 2 pyruvate Pyrococcus furiosus
-
 2-acetolactate + CO2
-
 ?
2.2.1.6 2 pyruvate Pyrococcus furiosus the enzyme produces 2-acetolactate in a temperature-dependent manner 2-acetolactate + CO2
-
 ?
2.2.1.6 additional information Pyrococcus furiosus non-enzymatic decarboxylation of acetolactate to acetoin ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.2.1.6 Pyrococcus furiosus Q8U2A4
-
-
2.2.1.6 Pyrococcus furiosus Q8U2A4 gene als
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.2.1.6 additional information no production of acetoin and butanediol at temperatures above 85°C, best production rates at 75-80°C, profile overview Pyrococcus furiosus
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.2.1.6 2 pyruvate
-
 Pyrococcus furiosus 2-acetolactate + CO2
-
 ?
2.2.1.6 2 pyruvate
-
 Pyrococcus furiosus 2-acetolactate + CO2 product decarboxylates spontaneously to final product acetoin (i.e. 3-hydroxybutanone), which is a major product at temperatures below 80°C. Acetolactate synthase ALS, which is involved in branched-chain amino acid biosynthesis, is responsible and deletion of the Als gene abolishes acetoin production ?
2.2.1.6 2 pyruvate the enzyme produces 2-acetolactate in a temperature-dependent manner Pyrococcus furiosus 2-acetolactate + CO2
-
 ?
2.2.1.6 2 pyruvate activated enzyme ALS catalyzes the condensation and decarboxylation of two pyruvates to one 2-acetolactate Pyrococcus furiosus 2-acetolactate + CO2
-
 ?
2.2.1.6 additional information non-enzymatic decarboxylation of acetolactate to acetoin Pyrococcus furiosus ?
-
 ?
2.2.1.6 additional information assay method with indirect quantitation of alpha-acetolactate by measuring acetoin formation Pyrococcus furiosus ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.2.1.6 acetolactate synthase
-
 Pyrococcus furiosus
2.2.1.6 ALS
-
 Pyrococcus furiosus
2.2.1.6 PF0935
-
 Pyrococcus furiosus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.2.1.6 75 78
-
 Pyrococcus furiosus
2.2.1.6 85
-
-
 Pyrococcus furiosus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.2.1.6 60 95 assay activity range Pyrococcus furiosus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.2.1.6 8.4
-
 assay at Pyrococcus furiosus

Cofactor

EC Number Cofactor Comment Organism Structure
2.2.1.6 FAD required Pyrococcus furiosus
2.2.1.6 thiamine diphosphate
-
 Pyrococcus furiosus
2.2.1.6 thiamine diphosphate dependent on Pyrococcus furiosus

Expression

EC Number Organism Comment Expression
2.2.1.6 Pyrococcus furiosus maximum gene expression between 72 and 80 °C additional information

General Information

EC Number General Information Comment Organism
2.2.1.6 malfunction deletion of the als gene abolishes acetoin production. Deletion of gene als in an engineered strain of Pyrococcus furiosus containing an additional pathway for ethanol production significantly improves the yield of ethanol Pyrococcus furiosus
2.2.1.6 malfunction deletion of the enzyme gene abolishes acetoin production Pyrococcus furiosus
2.2.1.6 metabolism there are two enzymes in nature that are known to produce 2-acetolactate from pyruvate, a catabolic ALS encoded by the alsSD operon, which is involved in the production of acetoin and butanediol, and ananabolic 2-acetohydroxyacid synthase (AHAS) that participates in biosynthesis of the branched chain amino acids isoleucine, leucine and valine. The flux of 2-acetolactate toward amino acid biosynthesis is regulated via allosteric activation of 2-acetolactate decarboxylase, ALDC, by either valine or leucine Pyrococcus furiosus
2.2.1.6 physiological function the enzyme is involved in the production of acetoin (3-hydroxybutanone) as a major product at growth temperatures below 80°C. Acetoin is produced by wild-type Pyrococcus furiosus during growth at sub-optimal temperatures below 80°C Pyrococcus furiosus
2.2.1.6 physiological function Pyrococcus furiosus produces acetoin as amajor end product at growth temperatures below 85°C in a temperature-dependent manner via a acetolactate synthase whose gene expression and biochemical function are temperature-dependent Pyrococcus furiosus