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Molecular characterization of Yeasts and Bacteria isolated from Handia, an Indian traditional rice fermented alcoholic beverage.

Joydeb Maji, Bidhan Chandra Mukhopadhyay, Suranjita Mitra, Swadesh Ranjan Biswas

Abstract


Handia is an Indian traditional fermented alcoholic beverage prepared from rice (Oryza sativa L.) To date, no information is available about the microorganisms associated with Handia fermentation. In this study a total of twelve yeast strains isolated from Handia were identified by partial sequencing of the D1/D2 variable domain of 26S of rRNA gene. The isolates were identified and designated as Saccharomyces cerevisiae  KpY,  S. cerevisiae 18VSL, S. cerevisiae H15, S. cerevisiae H17, Hanseniaspora guilliermondii G1,  H. guilliermondii G4,  Pichia kudriavzevii H21L, Candida glabrata H3, C. glabrata H8, C. glabrata H11, C. glabrata H12 and C. tropicalis 18VLL. Sequence analysis of 26S rRNA genes revealed 100% identity with the corresponding sequences in the GenBank with the exception of P. kudriavzevii H21L which showed 99% identity. The bacterial strains BA, B16 and B4 were identified as Brevibacillus agri, Leuconostoc mesenteroides and Kocuria sp., respectively by 16S rRNA gene sequence analysis. The identification of yeast strains (KpY, 18VSL, H15, H17, G4 and H21L) was further confirmed by 5.8S-ITS-RFLP analysis. M13 PCR-fingerprinting indicated the presence of four different strains of S. cerevisiae in Handia. The result suggests that Handia constitutes a complex microbial consortium comprising a wide diversity of bacterial and yeast strains.


Keywords


Handia; PCR-RFLP; ITS; fermentation; M13; diversity

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