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Abstract
The first development method of brewery shochu yeast focusing on chromosomal aneuploidy is reported in this study. Euploidy diploid shochu yeast S-3 was treated with a microtubule inhibitor, nocodazole, for the purpose of inducing aneuploidy. Next, 2,3,5-triphenyl tetrazolium chloride (TTC) staining and the growth rate were investigated to select aneuploids. Aneuploids were selected at a frequency of 8.2 × 10−4, which was significantly higher than that of the control group, mainly at chromosomes I, II, III, IX, XII, XIII, and XVI. The acquired aneuploids were evaluated for their metabolic and brewing characteristics. A hierarchical cluster analysis based on endogenous metabolite data discriminated euploid S-3 and aneuploids. In addition, principal-component analysis of the constituents of the broth brewed with the strains discriminated between euploid S-3 and aneuploids. Sensory evaluation of the broth brewed with euploid S-3 and aneuploids showed that it tended to differ in aroma and taste. Specific ethanol production rates of the aneuploids were not deteriorated. The method of this selection made it possible to efficiently obtain aneuploids with various brewing characteristics from brewer’s yeast, which do not correspond to genetically modified organisms. This novel breeding method focusing on chromosomal aneuploidy will facilitate the development of novel shochu yeast strains.
Highlights
It is made by distilling mash containing: koji; several kinds of crops such as barley, rice, and potato; and shochu yeast, Saccharomyces cerevisiae, which is used for ethanol fermentation
To develop a method to screen and obtain aneuploids from shochu yeasts, we focused on previous studies on nocodazole
To pioneer the possibility of developing a breeding method focused onto chromosomal aneuploidy in shochu yeast, we investigated the effect of nocodazole on the aneuploidy of shochu yeast and the brewery characteristics of the obtained aneuploids
Summary
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. A citric acid-producing fungus, Aspergillus luchuensis, was introduced in the early 1900s from the Okinawa islands, which made the mash acidic and prevented the proliferation of unfavorable microbes To cope with this revolution in the shochu industry, brewery yeasts tolerant to citric acid were isolated [1,2] in the same period and have been utilized until now. Numerous novel breeding strategies have been developed in sake yeasts [11], but breeding technologies for shochu yeasts have been developed These include protoplast fusion and isolating mutants tolerant to heat or NaCl to produce high-glycerol yeast [12,13]. To pioneer the possibility of developing a breeding method focused onto chromosomal aneuploidy in shochu yeast, we investigated the effect of nocodazole on the aneuploidy of shochu yeast and the brewery characteristics of the obtained aneuploids. We first succeeded in developing a method to induce chromosomal aneuploidy in shochu yeast and to alter its brewery characteristics
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