Molecular characterization of the Saccharomycopsis fibuligera ATF genes, encoding alcohol acetyltransferase for volatile acetate ester formation

Hye Yun Moon; Hyeon Jin Kim; Ki Seung Kim; Su Jin Yoo; Dong Wook Lee; Hee Je Shin; Jeong Ah Seo; Hyun Ah Kang

doi:10.1007/s12275-021-1159-8

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Molecular characterization of the Saccharomycopsis fibuligera ATF genes, encoding alcohol acetyltransferase for volatile acetate ester formation: Hye Yun Moon ¹, Hyeon Jin Kim ¹, Ki Seung Kim ¹, Su Jin Yoo ¹, Dong Wook Lee ¹, Hee Je Shin ¹, Jeong Ah Seo ², Hyun Ah Kang ¹; Journal of Microbiology 2021;59(6):598-608.
DOI: https://doi.org/10.1007/s12275-021-1159-8
Published online: May 29, 2021

¹Molecular Systems Biology Laboratory of Life Science, Chung-Ang University, Seoul 06974, Republic of Korea, ²School of Systems Biomedical Science, Soongsil University, Seoul 06978, Republic of Korea¹Molecular Systems Biology Laboratory of Life Science, Chung-Ang University, Seoul 06974, Republic of Korea, ²School of Systems Biomedical Science, Soongsil University, Seoul 06978, Republic of Korea

Corresponding author: Hyun Ah Kang , Tel: +82-2-820-5863,

Received: 29 March 2021 • Revised: 19 April 2021 • Accepted: 19 April 2021

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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.

Keywords: DNA physical analysis;Meiosis;Recombination;Saccharomyces cerevisiae;Southern blotting

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Molecular characterization of the Saccharomycopsis fibuligera ATF genes, encoding alcohol acetyltransferase for volatile acetate ester formation

J. Microbiol. 2021;59(6):598-608. Published online May 29, 2021

DOI: https://doi.org/10.1007/s12275-021-1159-8

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