This study reports the microbial quality of ethnic starter culture Xaj-pitha used for rice wine fermentation in Assam. Here, we collected 60 Xaj-pitha samples belonging to Ahom community of the state and enumerated the microorganisms using spread plate technique. Illumina-based whole genome shotgun sequencing detected the presence of microbial contaminants like Acidovorax, Herbaspirillum, Methylobacterium, Pantoea, Pseudomonas, Stenotrophomonas, Staphylococcus, Micrococcus, Acinetobacter, etc. Presence of major health hazards associated with spontaneous rice wine fermentation necessitated method optimization through the development of a defined mixed starter culture. For this, functionally important alpha-amylase producers viz., Penicillium sp. ABTSJ23, Rhizopus oryzae ABTSJ63, Mucor guilliermondii ABTSJ72 and Amylomyces rouxii ABTSJ82 and eight yeasts viz., Saccharomyces cerevisiae ABTY1J, ABTY1S, ADJ5 & ADJ1, Wickerhamomyces anomalus ADJ2, Saccharomycopsis malanga ADJ3, Saccharomycopsis fibuligera ADJ4 and Saccharomycopsis malanga ADJ6 were retrieved using appropriate media. All the mould cultures tested negative for aflotoxins production. Among the yeasts, Saccharomyces cerevisiae ABTY1S and ADJ1 decarboxylated lysine HCl and tyramine HCl, respectively, indicating their biogenic amine production ability. For defined mixed starter culture, Amylomyces rouxii ABT82 with alpha-amylase (5.92 U/ml) and glucoamylase (7.50 U/ml) activities was selected as fungal partner; while Saccharomycopsis fibuligera ADJ4 and Saccharomyces cerevisiae ABT-Y1J with high ethanol production (up to 10.11% and 9.88% v/v, respectively) were selected as yeast partners. The mixed culture was able to produce high amount of glucose, ethanol and liquid (glucose 10.91% w/v; ethanol 7.5% w/v; liquid 51.0% w/v). Therefore, this study demonstrated the efficiency of mixed starter cultures for safe and controlled rice wine production.
