Evolutionary engineering in chemostat cultures for improved maltotriose fermentation kinetics in saccharomyces pastorianus lager brewing yeast

Anja Brickwedde, Marcel van den Broek, Jan-Maarten A. Geertman, Frederico Magalhães, Niels G.A. Kuijpers, Brian Gibson, Jack T. Pronk, Jean-Marc G. Daran (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

30 Citations (Scopus)


The lager brewing yeast Saccharomyces pastorianus, an interspecies hybrid of S. eubayanus and S. cerevisiae, ferments maltotriose, maltose, sucrose, glucose and fructose in wort to ethanol and carbon dioxide. Complete and timely conversion ("attenuation") of maltotriose by industrial S. pastorianus strains is a key requirement for process intensification. This study explores a new evolutionary engineering strategy for improving maltotriose fermentation kinetics. Prolonged carbon-limited, anaerobic chemostat cultivation of the reference strain S. pastorianus CBS1483 on a maltotriose-enriched sugar mixture was used to select for spontaneous mutants with improved affinity for maltotriose. Evolved populations exhibited an up to 5-fold lower residual maltotriose concentration and a higher ethanol concentration than the parental strain. Uptake studies with 14C-labeled sugars revealed an up to 4.75-fold higher transport capacity for maltotriose in evolved strains. In laboratory batch cultures on wort, evolved strains showed improved attenuation and higher ethanol concentrations. These improvements were also observed in pilot fermentations at 1,000-L scale with high-gravity wort. Although the evolved strain exhibited multiple chromosomal copy number changes, analysis of beer made from pilot fermentations showed no negative effects on flavor compound profiles. These results demonstrate the potential of evolutionary engineering for strain improvement of hybrid, alloploid brewing strains.

Original languageEnglish
Article number1690
JournalFrontiers in Microbiology
Publication statusPublished - 8 Sept 2017
MoE publication typeA1 Journal article-refereed


This project is funded by the Seventh Framework Programme of the European Union in the frame of the SP3 people support for training and career development of researchers (Marie Curie), Networks for Initial Training (PITN-GA-2013 ITN-2013-606795) YeastCell. Brian Gibson was additionally supported by grant 276480 from the Academy of Finland. We thank Virve Vidgren for technical support with the transport assays.


  • brewing
  • chemostat
  • evolutionary engineering
  • maltose
  • maltotriose consumption rate
  • sacchromyces pastorianus
  • transport


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