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Food Microbiology

Enhanced Wort Fermentation with De Novo Lager Hybrids Adapted to High-Ethanol Environments

Kristoffer Krogerus, Sami Holmström, Brian Gibson
Donald W. Schaffner, Editor
Kristoffer Krogerus
VTT Technical Research Centre of Finland, Espoo, FinlandDepartment of Biotechnology and Chemical Technology, Aalto University, School of Chemical Technology, Aalto, Espoo, Finland
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Sami Holmström
VTT Technical Research Centre of Finland, Espoo, Finland
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Brian Gibson
VTT Technical Research Centre of Finland, Espoo, Finland
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Donald W. Schaffner
Rutgers, The State University of New Jersey
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DOI: 10.1128/AEM.02302-17
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ABSTRACT

Interspecific hybridization is a valuable tool for developing and improving brewing yeast in a number of industry-relevant aspects. However, the genomes of newly formed hybrids can be unstable. Here, we exploited this trait by adapting four brewing yeast strains, three of which were de novo interspecific lager hybrids with different ploidy levels, to high ethanol concentrations in an attempt to generate variant strains with improved fermentation performance in high-gravity wort. Through a batch fermentation-based adaptation process and selection based on a two-step screening process, we obtained eight variant strains which we compared to the wild-type strains in 2-liter-scale wort fermentations replicating industrial conditions. The results revealed that the adapted variants outperformed the strains from which they were derived, and the majority also possessed several desirable brewing-relevant traits, such as increased ester formation and ethanol tolerance, as well as decreased diacetyl formation. The variants obtained from the polyploid hybrids appeared to show greater improvements in fermentation performance than those derived from diploid strains. Interestingly, it was not only the hybrid strains, but also the Saccharomyces cerevisiae parent strain, that appeared to adapt and showed considerable changes in genome size. Genome sequencing and ploidy analysis revealed that changes had occurred at both the chromosome and single nucleotide levels in all variants. Our study demonstrates the possibility of improving de novo lager yeast hybrids through adaptive evolution by generating stable and superior variants that possess traits relevant to industrial lager beer fermentation.

IMPORTANCE Recent studies have shown that hybridization is a valuable tool for creating new and diverse strains of lager yeast. Adaptive evolution is another strain development tool that can be applied in order to improve upon desirable traits. Here, we apply adaptive evolution to newly created lager yeast hybrids by subjecting them to environments containing high ethanol levels. We isolated and characterized a number of adapted variants which possess improved fermentation properties and ethanol tolerance. Genome analysis revealed substantial changes in the variants compared to the original strains. These improved variant strains were produced without any genetic modification and are suitable for industrial lager beer fermentations.

FOOTNOTES

    • Received 18 October 2017.
    • Accepted 27 November 2017.
    • Accepted manuscript posted online 1 December 2017.
  • Supplemental material for this article may be found at https://doi.org/10.1128/AEM.02302-17.

  • Copyright © 2018 American Society for Microbiology.

All Rights Reserved.

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Enhanced Wort Fermentation with De Novo Lager Hybrids Adapted to High-Ethanol Environments
Kristoffer Krogerus, Sami Holmström, Brian Gibson
Appl. Environ. Microbiol. Jan 2018, 84 (4) e02302-17; DOI: 10.1128/AEM.02302-17

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Enhanced Wort Fermentation with De Novo Lager Hybrids Adapted to High-Ethanol Environments
Kristoffer Krogerus, Sami Holmström, Brian Gibson
Appl. Environ. Microbiol. Jan 2018, 84 (4) e02302-17; DOI: 10.1128/AEM.02302-17
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KEYWORDS

adaptive evolution
beer
ethanol
flavor
hybridization
yeast

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