SummaryWickerhamomyces anomalus D1‐4, a non‐Saccharomyces yeast known for its aroma production, offers a novel approach to improving the aroma and quality of mulberry wine. We employed various inoculation methods for fermenting mulberry wine and assessed its physicochemical, bioactive compounds, antioxidant activities, colour and aroma components. The results revealed that the addition of Wickerhamomyces anomalus D1‐4 to the fermentation led to a significant increase in the levels of alcohols, terpenes, nitrogen oxides and methyl in mulberry wine, especially, produced the characteristic isobutyl acetate and isopentyl hexanoate, and greatly increased the relative content of isopentyl acetate, isopentyl alcohol and phenylethyl alcohol. The sequential inoculation of Saccharomyces cerevisiae for 24 h followed by Wickerhamomyces anomalus D1‐4 fermented for 9 days was the best fermentation method for mulberry wine. Regarding colour, bioactive compounds and antioxidant activities, the fermentation was superior to other fermentations, and it was also able to maximise the aroma of mulberry wine and give it a strong floral and fruity aroma, especially higher ethyl hexanoate and ethyl octanoate. These findings demonstrate the unique fermentation characteristics of non‐Saccharomyces yeasts in enhancing the flavour and quality of mulberry wine.
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