Isoamyl acetate and ethyl caproate are the primary aroma compounds responsible for the fruity fragrance characteristic of Ginjo sake. Simultaneous high-level production of both compounds is crucial to achieving a balanced aroma and complex flavor. Isoamyl acetate is predominantly produced by hda1∆/hda1∆ and LEU4(G516S)/LEU4(G516S), while ethyl caproate is produced in high quantities by FAS2(G1250S)/FAS2(G1250S). In this study, to maximize the production of both aroma compounds, genome editing was employed to generate sake yeast strains combining these mutations. After small-scale fermentation tests were conducted to evaluate the production of aroma compounds, we found that the isoamyl acetate-enhancing effect of hda1∆/hda1∆ was almost completely masked by FAS2(G1250S)/FAS2(G1250S). In contrast, the effects of LEU4(G516S)/LEU4(G516S) were not entirely masked by FAS2(G1250S)/FAS2(G1250S), resulting in 2.4- and 5.4-fold greater production of isoamyl acetate and ethyl caproate, respectively. This study highlights the utility of genome editing in the combinatorial breeding of sake yeast.

• PMID: 40097305
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Gong Y, Klinkaewboonwong N, Hayashi R, Zhou Y, Nishida I, Saito R, Goshima T, Nishi T, Watanabe D, Hirata D, ... Show all

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