Water kefir is a sour, alcoholic, and fruity fermented beverage of which the fermentation is started with water kefir grains. These water kefir grains consist of polysaccharide and contain the microorganisms responsible for the water kefir fermentation. In this work, a water kefir fermentation process was followed as a function of time during 192 h to unravel the community dynamics, the species diversity, and the kinetics of substrate consumption and metabolite production. The majority of the water kefir ecosystem was found to be present on the water kefir grains. The most important microbial species present wereLactobacillus casei/paracasei,Lactobacillus harbinensis,Lactobacillus hilgardii,Bifidobacterium psychraerophilum/crudilactis,Saccharomyces cerevisiae, andDekkera bruxellensis. The microbial species diversities in the water kefir liquor and on the water kefir grains were similar and remained stable during the whole fermentation process. The major substrate, sucrose, was completely converted after 24 h of fermentation, which coincided with the production of the major part of the water kefir grain polysaccharide. The main metabolites of the fermentation were ethanol and lactic acid. Glycerol, acetic acid, and mannitol were produced in low concentrations. The major part of these metabolites was produced during the first 72 h of fermentation, during which the pH decreased from 4.26 to 3.45. The most prevalent volatile aroma compounds were ethyl acetate, isoamyl acetate, ethyl hexanoate, ethyl octanoate, and ethyl decanoate, which might be of significance with respect to the aroma of the end product.

Kombucha tea is made by aerobically fermenting a sweetened tea infusion with a kombucha culture, a symbiotic culture of bacteria and yeasts. The resulting beverage is usually non-alcoholic, sour and refreshing, but not naturally sparkling. Many consumers are drinking kombucha because of its alleged health effects and as kombucha tea is increasing in popularity, consumers are increasingly demanding kombucha beverages of better quality. To better adapt their products to these consumer demands, brewers are looking to the scientific knowledge about kombucha for answers. However, the scientific exploration of this complex naturally fermented beverage is still limited. This review paper presents the current state-of-the-art. To further deepen the understanding of the kombucha fermentation process and to help brewers in their search for higher-quality beverages, the microbial species diversity and its dynamics during the fermentation process should be investigated in detail, as well as the kinetics of the substrate consumption and metabolite production, and the relationship between the substrate and metabolite concentrations, and the consumer preferences for the resulting beverage.

A novel Bifidobacterium, strain LMG 28769T, was isolated from a household water kefir fermentation process. Cells were Gram-stain-positive, non-motile, non-spore-forming, catalase-negative, oxidase-negative and facultatively anaerobic short rods. Analysis of its 16S rRNA gene sequence revealed Bifidobacterium crudilactis and Bifidobacterium psychraerophilum (97.4 and 97.1 % similarity towards the respective type strain sequences) as nearest phylogenetic neighbours. Its assignment to the genus Bifidobacterium was confirmed by the presence of fructose 6-phosphate phosphoketolase activity. Analysis of the hsp60 gene sequence revealed very low similarity with nucleotide sequences in the NCBI nucleotide database. The genotypic and phenotypic analyses allowed the differentiation of strain LMG 28769T from all recognized Bifidobacterium species. Strain LMG 28769T ( = CCUG 67145T = R 54638T) therefore represents a novel species, for which the name Bifidobacterium aquikefiri sp. nov. is proposed.

Eight water kefir fermentation series differing in buffer capacity and calcium concentration of the water used for fermentation were studied during eight backslopping steps. High buffer capacities resulted in high pH values and high calcium concentrations resulted in low pH values at the end of each backslopping step. When the water buffer capacity and/or calcium concentration were below certain minima, the water kefir grain growth decreased gradually over multiple backsloppings. High water buffer capacities resulted in high concentrations of residual total carbohydrate concentrations and low metabolite concentrations. Further, high water buffer capacities resulted in high ratios of lactic acid bacteria to yeasts, which was reflected in high molar ratios of the concentrations of lactic acid to ethanol and acetic acid to ethanol. The most prevalent microorganisms of the water kefir grain inoculum and grains of all fermentation series at the end of the eighth backslopping step were Lactobacillus hilgardii, Lactobacillus nagelii, Lactobacillus paracasei, Bifidobacterium aquikefiri, Saccharomyces cerevisiae, and Dekkera bruxellensis. These microbial communities were influenced by the water buffer capacity and had an impact on the substrate consumption and metabolite production during water kefir fermentation.