Sugary Kefir: Microbial Identification and Biotechnological Properties

Background: The aim of the present study was to assess the microecosystem composition of three different fruit kefir grains used for the fermentation of apple juice (NAJ), cherry juice (SCN), and a solution of sugary water, enriched with plums (BSS). Methods: Yeast and bacterial populations were enumerated using classical microbiological techniques, clustered by RAPD-PCR genotyping, and identified by sequencing of the D1/D2 region of 26S-rRNA gene and the V1-V3 region of 16S-rRNA gene, respectively. The caseinolytic/lipolytic activities and the production of antimicrobial compounds were assessed by well diffusion assays. The proteolytic and lipolytic capacity were further assessed by SDS-PAGE and titrimetric assay, respectively. Results: Yeast and bacterial populations were enumerated at 6.28, 6.58, and 6.41 log CFU/g and at 4.32, 4.85, and 4.34 log CFU/g, regarding BSS, NAJ, and SCN, respectively. Saccharomyces cerevisiae dominated all three sugary kefir grains; Kluyveromyces marxianus formed a secondary microbiota in BSS and NAJ. Bacillus amyloliquefaciens dominated NAJ and SCN; Lactobacillus rhamnosus dominated BSS. Four bacteria and nine yeast isolates exhibited proteolytic activity. Forty bacteria and 45 yeast isolates possessed lipolytic activity. No antibacterial activity was detected. Conclusions: Prevalence of yeast over bacterial populations was evident in all samples assessed. Several bacterial and yeast strains exhibited proteolytic and lipolytic activities, making them suitable candidates for inclusion in starter cultures for milk and sugary kefir fermentation.