Evolutionary Engineering in Chemostat Cultures for Improved Maltotriose Fermentation Kinetics in Lager Brewing Yeast

Overview

Journal Front Microbiol

Specialty Microbiology

Date 2017 Sep 26

PMID 28943864

Citations 18

Authors

Anja Brickwedde

Marcel van den Broek

Jan-Maarten A Geertman

Frederico Magalhaes

Niels G A Kuijpers

Brian Gibson

Jack T Pronk

Jean-Marc G Daran

Affiliations

Soon will be listed here.

Abstract

The lager brewing yeast , an interspecies hybrid of and , ferments maltotriose, maltose, sucrose, glucose and fructose in wort to ethanol and carbon dioxide. Complete and timely conversion ("attenuation") of maltotriose by industrial 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 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 C-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.

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