Fermentation Process Optimization for High 2-Phenylethanol Aroma Whisky.

Jun 12, 2026International journal of molecular sciences

Improving Fermentation to Produce Whisky with Strong 2-Phenylethanol Aroma

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Abstract

The 2-phenylethanol (2-PE) content in whisky fermented by the modified yeast reached 3.68 g/L, a significant increase.

The typical yield of 2-PE in brewed wines is less than 100 mg/L.
Gene editing using CRISPR-Cas9 was employed to delete the gene encoding aromatic transaminase I in yeast.
Fermentation conditions were optimized, resulting in a 23.73% increase in 2-PE content compared to the original yeast strain.
Optimal fermentation conditions included a sugar concentration of 46.30 g/L, a fermentation time of 6 days, and an L-phenylalanine concentration of 1.43 g/L.
This approach could enhance the flavor profile of whisky and other brewed wines.

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2-Phenylethanol (2-PE) is a key aromatic alcohol contributing to the rose-like odor in brewed wines, primarily synthesized by yeast metabolism with a typical yield of less than 100 mg/L. To enhance the 2-PE content in brewed wines, this study used CRISPR-Cas9 gene editing technology to delete thegene (encoding aromatic transaminase I) inSY. The single-factor experiments were performed to optimize the fermentation process, and the 2-PE content in the brewed wine was measured by high-performance liquid chromatography. The results demonstrated that the 2-PE content in whisky fermented by the SY-A8 was 0.73 g/L, increasing 23.73% compared to SY. The fermentation conditions of SY-A8 were optimized through single-factor experiments and the Box-Behnken design. The optimal conditions were a sugar concentration of 46.30 g/L, a fermentation time of 6 days, and an L-phenylalanine concentration of 1.43 g/L. The high 2-phenylethanol aroma whisky was brewed with a higher 2-phenylethanol content of 3.68 g/L in a 1 L fermenter at the optimal conditions. In conclusion, the modification ofby CRISPR-Cas9 gene editing combined with fermentation process optimization provides an effective technical strategy for improving the 2-PE content in whisky, thereby providing a research perspective for the flavor enhancement of whisky and other brewed wines. ARO8Saccharomyces cerevisiae Saccharomyces cerevisiae

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Fermentation Process Optimization for High 2-Phenylethanol Aroma Whisky.

Abstract

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