Isolation, Biochemical Characterization and DNA Sequencing of Microorganisms from Fermented Kanji

Abstract

Kanji is a traditional fermented beverage prepared from beetroot (Beta vulgaris), black carrot (Daucus carota), and mustard seeds (Brassica juncea), valued for its probiotic and functional properties. In this study, kanji was prepared under laboratory conditions using raw materials collected from the Dehradun market and fermented at room temperature for 21 days. During fermentation, the pH of kanji typically decreases from approximately 5.3 to 3.3, largely due to the metabolic activity of bacteria such as Lactic acid bacteria, and this acidification creates an inhibitory environment that suppresses the growth of pathogenic microorganisms, thereby enhancing the microbial safety of the final product.  Physio-chemical parameters including colour, taste, pH, and total titratable acidity were monitored at regular intervals, revealing a gradual decrease in pH and increased acidity due to lactic acid fermentation. Microbial analysis was performed using the standard plate count method, which showed lactic acid bacteria (LAB) as the dominant microbial group. LAB were isolated using serial dilution and MRS agar, followed by purification and biochemical characterization. Selective media were employed to detect potential pathogenic bacteria. Molecular identification of dominant LAB isolates were carried out using 16S rRNA gene sequencing to ensure accurate taxonomic resolution. The combined microbiological, biochemical, and molecular approach provided comprehensive insights into the microbial ecology of fermented kanji and supported its potential as a safe, nutrient rich probiotics, immunity booster, functional fermented beverage