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Application of directed evolution to develop ethanol tolerant Oenococcus oeni for more efficient malolactic

Alice L Betteridge1, Krista M Sumby1, Joanna F Sundstrom1

  • 1Department of Wine and Food Science, School of Agriculture, Food and Wine, University of Adelaide, Waite Campus, Urrbrae, South Australia, Australia.

Applied Microbiology and Biotechnology
|November 19, 2017
PubMed
Summary

Directed evolution created a robust Oenococcus oeni strain for winemaking. The evolved O. oeni (A90) shows improved malolactic fermentation performance in high ethanol conditions.

Keywords:
Directed evolutionEthanolMalolactic fermentationOenococcusStress responseWine

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Area of Science:

  • Microbiology
  • Enology
  • Biotechnology

Background:

  • Malolactic fermentation (MLF) is crucial in winemaking but often unreliable due to Oenococcus oeni's sensitivity.
  • Oenococcus oeni's fastidious nature limits its application in challenging wine environments.

Purpose of the Study:

  • To improve Oenococcus oeni robustness for winemaking using directed evolution.
  • To develop a strain capable of withstanding high ethanol concentrations during MLF.

Main Methods:

  • Established a continuous culture of O. oeni exposed to gradually increasing ethanol levels over approximately 330 generations.
  • Assessed the fermentation performance and ethanol tolerance of an evolved isolate (A90).
  • Compared gene expression profiles, specifically hsp18, between the evolved strain (A90) and the parent strain (SB3).

Main Results:

  • An evolved O. oeni isolate (A90) completed MLF faster in high ethanol medium compared to the parent strain.
  • Strain A90 demonstrated enhanced ethanol tolerance and improved fermentation performance in wine.
  • Increased hsp18 gene expression was observed in A90 under high ethanol stress, indicating a role in tolerance.

Conclusions:

  • Directed evolution is a successful strategy for improving Oenococcus oeni strains for winemaking.
  • The study provides insights into the genetic basis of ethanol tolerance in O. oeni.
  • The evolved strain A90 represents a promising candidate for industrial wine production.