Diacetyl control during brewery fermentation via adaptive laboratory engineering of the lager yeast Saccharomyces pastorianus

Brian Gibson (Corresponding Author), Virve Vidgren, Gopal Peddinti, Kristoffer Krogerus

    Research output: Contribution to journalArticleScientificpeer-review

    29 Citations (Scopus)


    Diacetyl contributes to the flavor profile of many fermented products. Its typical buttery flavor is considered as an off flavor in lager-style beers, and its removal has a major impact on time and energy expenditure in breweries. Here, we investigated the possibility of lowering beer diacetyl levels through evolutionary engineering of lager yeast for altered synthesis of α-acetolactate, the precursor of diacetyl. Cells were exposed repeatedly to a sub-lethal level of chlorsulfuron, which inhibits the acetohydroxy acid synthase responsible for α-acetolactate production. Initial screening of 7 adapted isolates showed a lower level of diacetyl during wort fermentation and no apparent negative influence on fermentation rate or alcohol yield. Pilot-scale fermentation was carried out with one isolate and results confirmed the positive effect of chlorsulfuron adaptation. Diacetyl levels were over 60% lower at the end of primary fermentation relative to the non-adapted lager yeast and no significant change in fermentation performance or volatile flavor profile was observed due to the adaptation. Whole-genome sequencing revealed a non-synonymous SNP in the ILV2 gene of the adapted isolate. This mutation is known to confer general tolerance to sulfonylurea compounds, and is the most likely cause of the improved tolerance. Adaptive laboratory evolution appears to be a natural, simple and cost-effective strategy for diacetyl control in brewing.
    Original languageEnglish
    Pages (from-to)1103-1112
    JournalJournal of industrial microbiology and biotechnology
    Issue number12
    Publication statusPublished - 1 Dec 2018
    MoE publication typeA1 Journal article-refereed


    Acknowledgements Open access funding provided by Technical Research Centre of Finland (VTT). This work was supported by PBL Brewing Laboratory (Oy Panimolaboratorio—Bryggerilaboratorium Ab), Tekes, the Finnish Funding Agency for Technology and Innovation, the Alfred Kordelin Foundation, Svenska Kulturfonden—The Swedish Cultural Foundation in Finland, Suomen Kulttuurirahasto, and the Academy of Finland (Academy Project 276480).


    • Beer
    • Chlorsulfuron
    • Diacetyl
    • Saccharomyces pastorianus
    • α-Acetolactate


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