Breeding of high-tolerance yeast by adaptive evolution and high gravity brewing of mutant.

Abstract

Ethanol and osmotic stresses are the major limiting factors for brewing strong-beer with high-gravity wort. Breeding of yeast strains with high osmotic and ethanol tolerance and studying very high gravity (VHG) brewing technology is of great significance for brewing strong-beer. This study used an optimized microbial microdroplet culture (MMC) system for adaptive laboratory evolution (ALE) of Saccharomyces cerevisiae YN81 to improve its tolerance to osmotic and ethanol stress. Meanwhile, we investigated the VHG and VHG with added ethanol (VHGAE) brewing processes for the evolved mutants in brewing strong-beer. The results showed that three evolved mutants were obtained; among them, the growth performance of YN81mc-8.3 under 300, 340, 380, 420, and 460 g·L-1 sucrose stresses was greater than that of the other strains. The ethanol tolerance of YN81mc-8.3 was 12%, which was 20% higher than that of YN81. During strong-beer brewing in a 100 L cylindrical cone-bottom tank, the sugar utilization and ethanol yield of YN81mc-8.3 outperformed those of YN81 in both the VHG and VHGAE brewing processes. Measurement of the diacetyl concentration showed that YN81mc-8.3 had a stronger diacetyl reduction ability; in particular, the real degree of fermentation of beers brewed by YN81mc-8.3 in VHG and VHGAE brewing processes was 75.35% and 66.71%, respectively, higher than those of the 2 samples brewed by YN81. Meanwhile, the visual, olfactive, and gustative properties of the strong-beer produced by YN81mc-8.3 were better than those of the other beers. In this study, the mutant YN81mc-8.3 and the VHGAE brewing process were optimal and represented a better alternative strong-beer brewing process. This article is protected by copyright. All rights reserved.