Affiliation:
1. grid.416629.e 0000 0004 0377 2137 Research Institute, Gekkeikan Sake Co. Ltd. 101 Shimotoba-koyanagi-cho 612-8385 Fushimi-ku Kyoto Japan
2. grid.260493.a 0000 0000 9227 2257 Graduate School of Science and Technology Nara Institute of Science and Technology 8916-5 Takayama 630-0192 Ikoma Nara Japan
Abstract
Abstract
Sake is a traditional Japanese alcoholic beverage brewed with the yeast Saccharomyces cerevisiae. Sake taste is affected by sugars, organic acids, and amino acids. We previously isolated mutants resistant to the proline analogue azetidine-2-carboxylate derived from a diploid sake yeast strain. Some of the mutants produced a greater amount of proline in the brewed sake. One of them (strain K-9-AZC) carried a novel mutation in the PRO1 gene encoding the Gln79His variant of the γ-glutamyl kinase Pro1, a key enzyme in proline biosynthesis in S. cerevisiae. This mutation resulted in extreme desensitization to feedback inhibition by proline, leading to proline overproduction. Interestingly, sake brewed with K-9-AZC contained 3.7-fold more proline, but only 25% less succinate than sake brewed with the parent strain. Metabolome analysis suggests that the decrease in succinate was attributable to a lower level of 2-oxoglutarate, which is converted into glutamate. The approach here could be a practical method for breeding of yeast strains involved in the diversity of sake taste.
Funder
The NARO Bio-oriented Technology Research Advancement Institution
Publisher
Oxford University Press (OUP)
Subject
Applied Microbiology and Biotechnology,Biotechnology,Bioengineering
Cited by
8 articles.
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