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Literature summary extracted from

  • Loebs, A.K.; Schwartz, C.; Thorwall, S.; Wheeldon, I.
    Highly multiplexed CRISPRi repression of respiratory functions enhances mitochondrial localized ethyl acetate biosynthesis in Kluyveromyces marxianus (2018), ACS Synth. Biol., 7, 2647-2655 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.268 gene EAT1, recombinant expression of N-terminally truncated mutants Eat1(DELTA1-14) and Eat1(DELTA1-19) in Saccharomyces cerevisiae and Kluyveromyces marxianus, recombinant expression of GFP-tagged wild-type enzyme in Kluyveromyces marxianus Kluyveromyces marxianus

Protein Variants

EC Number Protein Variants Comment Organism
2.3.1.268 additional information construction of an Eat1 knockout mutant and of N-terminally truncated mutants. Modulation of expression of the TCA cycle and electron transport chain genes using a highly multiplexed CRISPRi approach, simultaneous knockdown of ACO2b, SDH2, RIP1, and MSS51 results in a 3.8fold increase in ethyl acetate productivity over the already high natural capacity. Mitochondrial localization is partially lost when the first 14 amino acids are removed with complete cytosolic expression resulting from truncation at the predicted cut site, i.e. mutants Eat1(DELTA1-14) and Eat1(DELTA1-19) Kluyveromyces marxianus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.3.1.268 mitochondrion the N-terminal domain of enzyme Eat1 is responsible for targeting Kluyveromyces marxianus 5739
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.1.268 ethanol + acetyl-CoA Kluyveromyces marxianus
-
 ethyl acetate + CoA
-
 ?
2.3.1.268 ethanol + acetyl-CoA Kluyveromyces marxianus NBRC 104275
-
 ethyl acetate + CoA
-
 ?
2.3.1.268 ethanol + acetyl-CoA Kluyveromyces marxianus BCC 29191
-
 ethyl acetate + CoA
-
 ?
2.3.1.268 ethanol + acetyl-CoA Kluyveromyces marxianus DMKU3-1042
-
 ethyl acetate + CoA
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.268 Kluyveromyces marxianus W0T4A7
-
-
2.3.1.268 Kluyveromyces marxianus BCC 29191 W0T4A7
-
-
2.3.1.268 Kluyveromyces marxianus DMKU3-1042 W0T4A7
-
-
2.3.1.268 Kluyveromyces marxianus NBRC 104275 W0T4A7
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.268 ethanol + acetyl-CoA
-
 Kluyveromyces marxianus ethyl acetate + CoA
-
 ?
2.3.1.268 ethanol + acetyl-CoA
-
 Kluyveromyces marxianus NBRC 104275 ethyl acetate + CoA
-
 ?
2.3.1.268 ethanol + acetyl-CoA
-
 Kluyveromyces marxianus BCC 29191 ethyl acetate + CoA
-
 ?
2.3.1.268 ethanol + acetyl-CoA
-
 Kluyveromyces marxianus DMKU3-1042 ethyl acetate + CoA
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.1.268 alcohol acetyltransferase
-
 Kluyveromyces marxianus
2.3.1.268 eat1
-
 Kluyveromyces marxianus

Cofactor

EC Number Cofactor Comment Organism Structure
2.3.1.268 acetyl-CoA
-
 Kluyveromyces marxianus

General Information

EC Number General Information Comment Organism
2.3.1.268 malfunction modulation of expression of the TCA cycle and electron transport chain genes using a highly multiplexed CRISPRi approach, simultaneous knockdown of ACO2b, SDH2, RIP1, and MSS51 results in a 3.8fold increase in ethyl acetate productivity over the already high natural capacity Kluyveromyces marxianus
2.3.1.268 metabolism the enzyme is involved in the the primary pathways of precursor and acetate ester biosynthesis Kluyveromyces marxianus
2.3.1.268 physiological function the alcohol acetyltransferase Eat1 is the critical enzyme for ethyl, isoamyl, and phenylethyl acetate production. Ethyl acetate is formed from the condensation of ethanol and acetyl-CoA. Mitochondrial localization of Eat1 is necessary for high ethyl acetate production Kluyveromyces marxianus