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Background: Cayratia trifolia has been extensively studied for its bioactive components and medicinal properties. This study was carried out to evaluate the fermentation ability of Saccharomyces cerevisiae 2.1 yeast to determine suitable fermentation conditions. Methods: The initial sugar content for three-leaf cayratia fermentation and fermentation efficiency were calculated. The temperature for three-leaf cayratia fermentation, incubation time, and preliminary 1 liter of three-leaf cayratia wine fermentation were determined. All treatments showed that the total sugar content decreased after 9 days of fermentation compared to the initial level. Temperature during fermentation has a direct effect on yeast activity. Temperature increases the growth of yeast and speed of enzyme activity. The fermentation time changes depending on temperature, initial soluble solid contents, and yeast strains, leading to changes in the ethanol content after the end of fermentation. Results: The results showed that the S. cerevisiae 2.1 strain was able to ferment at room temperature with an initial pH of 4.5, 24 °Brix, and a yeast density of 107 cells/mL. The appropriate fermentation conditions determined for S. cerevisiae 2.1 yeast were 24 °Brix and incubation at room temperature (28-33°C) with a fermentation time of 11 days. Concusion: In 1 liter scale fermentation, three-leaf cayratia wine had 9.46% (v/v) and the fermentation efficiency was 90.34%. The wine produced had a unique color, flavor, and aroma that met the sensory evaluation of the Vietnamese standard TCVN-3217:79.
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