Isolation and Identification of Yeasts in Marcha, a Rice Wine Starter Culture From Nepal

Nepal harbors a rich diversity of cultures and traditions, including the unique practice of creating an indigenous grain called Marcha by various ethnic groups such as Newar, Tamang, Sherpa, Rai, Limbu, Gurung, Magar, and Tharu people. In the eastern region of Nepal, Marcha producers utilize over 42 different plants, including Vernonia cinerea, Clematis grewiae, Polygala arillata, Buddleja asiatica, Inula sp., Scoparia, and more, which shows regional diversity. The primary objective of the study was to explore the diversity of yeast present in Marcha samples. The studied Marcha samples were collected from 10 different geographic regions of Nepal, which included altogether 27 samples. The isolates were grouped into Groups A, B, and C based on morphological and physiological characteristics. Notably, Group B yeast displayed high amylase production, an enzyme responsible for starch breakdown, and exhibited the ability to produce ethanol. To further investigate the potential of these isolates, stress exclusion tests were conducted, with 30 isolates (70%) showing positive responses. The yeast isolates demonstrated resilience to high glucose concentrations of up to 36% (w/v) at a pH above 3 and a temperature of 37°C, which is the ideal growth condition. The study observed a direct correlation between the yeast isolates' ethanol production capabilities and their tolerance to different ethanol concentrations. Considering that all tested Marcha samples contained yeast capable of starch degradation and ethanol production, it was expected that these yeast isolates would actively participate in the fermentation of starch-based alcohol.