title: Ethyl acetate production by the elusive alcohol acetyltransferase from yeast.
abstract: Ethyl acetate is an industrially relevant ester that is currently produced exclusively through unsustainable processes. Many yeasts are able to produce ethyl acetate, but the main responsible enzyme has remained elusive, hampering the engineering of novel production strains. Here we describe the discovery of a new enzyme (Eat1) from the yeast Wickerhamomyces anomalus that resulted in high ethyl acetate production when expressed in Saccharomyces cerevisiae and Escherichia coli. Purified Eat1 showed alcohol acetyltransferase activity with ethanol and acetyl-CoA. Homologs of eat1 are responsible for most ethyl acetate synthesis in known ethyl acetate-producing yeasts, including S. cerevisiae, and are only distantly related to known alcohol acetyltransferases. Eat1 is therefore proposed to compose a novel alcohol acetyltransferase family within the α/β hydrolase superfamily. The discovery of this novel enzyme family is a crucial step towards the development of biobased ethyl acetate production and will also help in selecting improved S. cerevisiae brewing strains.
authors.affiliation: Laboratory of Microbiology, Wageningen University and Research, Stippeneng 4, 6708 WE Wageningen, The Netherlands. Electronic address: alex.kruis@wur.nl.
authors.forename: Aleksander J
authors.initials: AJ
authors.lastname: Kruis
authors.affiliation: Laboratory of Microbiology, Wageningen University and Research, Stippeneng 4, 6708 WE Wageningen, The Netherlands. Electronic address: mark.levisson@wur.nl.
authors.forename: Mark
authors.initials: M
authors.lastname: Levisson
authors.affiliation: Biobased Products, Wageningen University and Research,Bornse Weilanden 9, 6708 WG Wageningen, The Netherlands. Electronic address: astrid.mars@wur.nl.
authors.forename: Astrid E
authors.initials: AE
authors.lastname: Mars
authors.affiliation: Biobased Products, Wageningen University and Research,Bornse Weilanden 9, 6708 WG Wageningen, The Netherlands.
authors.forename: Max
authors.initials: M
authors.lastname: van der Ploeg
authors.affiliation: Laboratory of Microbiology, Wageningen University and Research, Stippeneng 4, 6708 WE Wageningen, The Netherlands.
authors.forename: Fernando
authors.initials: F
authors.lastname: Garcés Daza
authors.affiliation: Laboratory of Microbiology, Wageningen University and Research, Stippeneng 4, 6708 WE Wageningen, The Netherlands.
authors.forename: Valeria
authors.initials: V
authors.lastname: Ellena
authors.affiliation: Laboratory of Microbiology, Wageningen University and Research, Stippeneng 4, 6708 WE Wageningen, The Netherlands. Electronic address: serve.kengen@wur.nl.
authors.forename: Servé W M
authors.initials: SWM
authors.lastname: Kengen
authors.affiliation: Laboratory of Microbiology, Wageningen University and Research, Stippeneng 4, 6708 WE Wageningen, The Netherlands. Electronic address: john.vanderoost@wur.nl.
authors.forename: John
authors.initials: J
authors.lastname: van der Oost
authors.affiliation: Bioprocess Engineering, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands. Electronic address: ruud.weusthuis@wur.nl.
authors.forename: Ruud A
authors.initials: RA
authors.lastname: Weusthuis
mesh_terms: D000085:Acetates / Q000378:metabolism*; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
chemical_list: D000085:Acetates; D005656:Fungal Proteins; D011506:Proteins; C007650:ethyl acetate; C079238:alcohol O-acetyltransferase
keywords: Alcohol acetyltransferase; Escherichia coli; Ethyl acetate; Saccharomyces cerevisiae; yeast; α/β hydrolase