The current study investigated the in vitro probiotic potential of yeast isolated from kombucha, a tea beverage fermented with a symbiotic culture of acetic acid bacteria and yeast. A total of 62 yeast strains were previously isolated from four different commercial kombucha samples sold in New Zealand. Fifteen representative isolates belonging to eight different species were evaluated for their growth under different conditions (temperature, low pH, concentrations of bile salts, and NaCl). Cell surface characteristics, functional and enzymatic activities of the selected strains were also studied in triplicate experiments. Results showed that six strains (Dekkera bruxellensis LBY1, Sachizosaccharomyces pombe LBY5, Hanseniaspora valbyensis DOY1, Brettanomyces anomalus DOY8, Pichia kudraivzevii GBY1, and Saccharomyces cerevisiae GBY2) were able to grow under low-acid conditions (at pH 2 and pH 3) and in the presence of bile salts. This suggests their potential to survive passage through the human gut. All 15 strains exhibited negative enzymatic activity reactions (haemolytic, gelatinase, phospholipase, and protease activities), and thus, they can be considered safe to consume. Notably, two of the fifteen strains (Pichia kudraivzevii GBY1 and Saccharomyces cerevisiae GBY2) exhibited desirable cell surface hydrophobicity (64.60–83.87%), auto-aggregation (>98%), co-aggregation, resistance to eight tested antibiotics (ampicillin, chloramphenicol, colistin sulphate, kanamycin, nalidixic acid, nitrofurantoin, streptomycin, and tetracycline), and high levels of antioxidant activities (>90%). Together, our data reveal the probiotic activities of two yeast strains GBY1 and GBY2 and their potential application in functional food production.

Kombucha is a fermented tea beverage containing live microorganisms, mostly beneficial strains of yeast and acetic acid bacteria (AAB), but empirical evidence is limited supporting the probiotic potential of kombucha. This study reports the in vitro probiotic potential of 36 AAB strains isolated from three Kombucha samples commercially available in New Zealand. Nine representative AAB strains, belonging to three species (Komagataeibacter rhaeticus, Acetobacter musti and Gluconobacter potus), were examined for their primary probiotic characteristics such as tolerance to bile salts, NaCl, and low pH, and temperature. Three non-cellulose forming strains (A. musti LOAAB1, G. potus LOAAB2 and G. potus GBAAB3) were assayed for their cell surface characteristics such as auto-aggregation, co-aggregation with pathogenic bacteria, and hydrophobicity. Antimicrobial, antioxidant activities and enzymatic activities were also investigated for the strains of interest. Results indicated that nine strains were able to grow under low pH in the presence of bile salts, suggesting their potential to survive in the human gut. Six K. rhaeticus strains produced cellulosic pellicles, a potential source of prebiotics for beneficial bacteria. The three AAB strains LOAAB1, LOAAB2 and GBAAB3 showed promising cell surface characteristics, such as auto-aggregation rates (>80 %), co-aggregation with four pathogenic bacteria (13.24–43.47 %), hydrophobicity (42.12 % to 50.20 %), and antioxidant activities (>90 %). All nine strains tested negative for enzymatic activities (haemolytic, proteolytic, phospholipase, and gelatinase), suggesting that they are safe to consume. Together, these data indicate the potential for the three AAB strains to be further investigated as probiotic sources with more in vivo tests for applications in the food and beverages industry.

Kombucha is a popular sparkling sugared tea, fermented by a symbiotic culture of acetic acid bacteria (AAB) and yeast. The demand for kombucha continues to increase worldwide, mainly due to its perceived health benefits and appealing sensory properties. This study isolated and characterised the dominant AAB and yeast from a starter culture and kombucha broth after 0, 1, 3, 5, 7, 9, 11, and 14 days of fermentation at ambient temperature (22 °C). Yeast and AAB were isolated from the Kombucha samples using glucose yeast extract mannitol ethanol acetic acid (GYMEA) and yeast extract glucose chloramphenicol (YGC) media, respectively. The phenotypic and taxonomic identification of AAB and yeast were determined by morphological and biochemical characterisation, followed by a sequence analysis of the ribosomal RNA gene (16S rRNA for AAB and ITS for yeast). The changes in the microbial composition were associated with variations in the physico-chemical characteristics of kombucha tea, such as pH, titratable acidity, and total soluble solids (TSS). During fermentation, the acidity increased and the TSS decreased. The yield, moisture content, and water activity of the cellulosic pellicles which had developed at the end of fermentation were attributed to the presence of AAB. The dominant AAB species in the cellulosic pellicles and kombucha broth were identified as Komagataeibacter rhaeticus. The yeast isolates belonged to Debaryomyces prosopidis and Zygosaccharomyces lentus.

The demand for Kombucha, a sparkling sugared tea beverage fermented by a symbiotic culture of acetic acid bacteria (AAB) and yeast is increasing worldwide. Despite the popularity of the beverage which is mainly due to its perceived health benefits and appealing sensory properties, the microbial composition of the products at the time of consumption is unknown. Such information is important to both manufacturers and consumers. Therefore, this study characterised the dominant AAB and yeast present in six commercial Kombucha samples sold in New Zealand which comprised of three domestic and three imported samples. Acetic acid bacteria and yeast were isolated from the Kombucha samples using glucose yeast extract peptone mannitol (GYPM) and yeast extract glucose chloramphenicol (YGC) media, respectively. Phenotypic and taxonomic identification of AAB and yeast were achieved by morphological and biochemical characterisation, followed by sequence analysis of ribosomal RNA genes (16S rRNA for AAB and 26S rRNA for yeast). Viable AAB and yeast were only found in domestically produced Kombucha samples and not in the imported products. The dominant AAB species were identified as Acetobacter musti and Gluconobacter potus. The yeast isolates belonged to Dekkera bruxelensis, Schizosaccharomyces pombes, Hanseniaspora valbyensis, Brettanomyces anamalus, Pichia kudriavzevii, Starmerella vitis and Saccharomyces cerevisiae. The yeast communities were more complex and variable than the AAB communities in the analysed Kombucha samples.

Kombucha is a sparkling sugared tea commonly prepared using a sugared tea infusion and fermented at ambient temperature for several days using a cellulose pellicle also called tea fungus that is comprised of acetic acid bacteria and yeast. Consumption of Kombucha has been reported as early as 220 B.C. with various reported potential health benefits and appealing sensory properties. During Kombucha fermentation, sucrose is hydrolysed by yeast cells into fructose and glucose, which are then metabolised to ethanol. The ethanol is then oxidised by acetic acid bacteria (AAB) to produce acetic acid which is responsible for the reduction of the pH and also contributes to the sour taste of Kombucha. Characterisation of the AAB and yeast in the Kombucha starter culture can provide a better understanding of the fermentation process. This knowledge can potentially aid in the production of higher quality products as these microorganisms affect the production of metabolites such as organic acids which are associated with potential health benefits, as well as sensory properties. This review presents recent advances in the isolation, enumeration, biochemical characteristics, conventional phenotypic identification system, and modern genetic identification techniques of AAB and yeast present in Kombucha to gain a better understanding of the microbial diversity of the beverage.