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Abstract
Sake yeast is mostly diploid, so the introduction of recessive mutations to improve brewing characteristics requires considerable effort. To construct sake yeast with multiple excellent brewing characteristics, we used an evidence-based approach that exploits genome editing technology. Our breeding targeted the AWA1, CAR1, MDE1, and FAS2 genes. We introduced eight mutations into standard sake yeast to construct a non-foam-forming strain that makes sake without producing carcinogens or an unpleasant odor, while producing a sweet ginjo aroma. Small-scale fermentation tests showed that the desired sake could be brewed with our genome-edited strains. The existence of a few unexpected genetic perturbations introduced during breeding proved that genome editing technology is extremely effective for the serial breeding of sake yeast.
Highlights
Sake is an alcoholic beverage that originated in Japan and has been made for centuries
Based on whole-genome sequencing and morphological analyses of the genome-edited strains, we propose that genome editing technology can be applied for the effective construction of sake yeast strains by serial breeding
By using any of the genome-edited sake yeasts created in this study, the effective capacity of a brewing tank can be increased because awa1∆/awa1∆ possesses a non-foam-forming characteristic
Summary
Sake is an alcoholic beverage that originated in Japan and has been made for centuries. It is made by parallel double fermentation of sake rice with a high starch content, mainly by the action of two types of microorganisms [1]. Sake yeast originated in Japan and is tolerant to high alcohol concentration and low pH, with a high fermentation ability for making sake with excellent flavor characteristics. These characteristics indicate that sake yeast has been domesticated for a unique purpose in a geographically isolated situation
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