Abstract
Aroma ester components produced by fermenting yeast cells
via alcohol acetyltransferase (AATase)-catalyzed intracellular
reactions are responsible for the fruity character of fermented
alcoholic beverages, such as beer and wine. Acetate esters
are reportedly produced at relatively high concentrations by
non-Saccharomyces species. Here, we identified 12 ATF orthologues
(SfATFs) encoding putative AATases, in the diploid
genome of Saccharomycopsis fibuligera KJJ81, an isolate from
wheat-based Nuruk in Korea. The identified SfATF proteins
(SfAtfp) display low sequence identities with S. cerevisiae
Atf1p (between 13.3 and 27.0%). All SfAtfp identified, except
SfAtf(A)4p and SfAtf(B)4p, contained the activation domain
(HXXXD) conserved in other Atf proteins. Culture supernatant
analysis using headspace gas chromatography mass spectrometry
confirmed that the recombinant S. cerevisiae strains
expressing SfAtf(A)2p, SfAtf(B)2p, and SfAtf(B)6p produced
high levels of isoamyl and phenethyl acetates. The volatile
aroma profiles generated by the SfAtf proteins were distinctive
from that of S. cerevisiae Atf1p, implying difference in
the substrate preference. Cellular localization analysis using
GFP fusion revealed the localization of SfAtf proteins proximal
to the lipid particles, consistent with the presence of amphipathic
helices at their N- and C-termini. This is the first
report that systematically characterizes the S. fibuligera ATF
genes encoding functional AATases responsible for acetate
ester formation using higher alcohols as substrate, demonstrating
their biotechnological potential for volatile ester production.
Citations
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