The use of yeast assist kefir fermentation, but also can cause food spoilage if uncontrolled. Hence, in this study, the microbial composition of an existing commercial kefir starter was modified to produce a functional starter, where Lactobacillus acidophilus KCNU and Lactobacillus brevis Bmb6 were used to replace yeast in the original starter to produce non-yeast kefir-like fermented milk. The functional starter containing L. acidophilus KCNU and L. brevis Bmb6 demonstrated excellent stability with 10 10 CFU/g of total viable cells throughout the 12 weeks low-temperature storage. The newly developed functional starter also displayed a similar fermentation efficacy as the yeast-containing control starter, by completing the milk fermentation within 12 h, with a comparable total number of viable cells (10 8 CFU/mL) in the final products, as in control. Sensory evaluation revealed that the functional starterfermented milk highly resembled the flavor of the control kefir, with enhanced sourness. Furthermore, oral administration of functional starter-fermented milk significantly improved the disease activity index score by preventing drastic weight-loss and further deterioration of disease symptoms in DSS-induced mice. Altogether, L. acidophilus KCNU and L. brevis Bmb6 have successfully replaced yeast in a commercial starter pack to produce a kefir-like fermented milk beverage with additional health benefits. The outcome of this study provides an insight that the specific role of yeast in the fermentation process could be replaced with suitable probiotic candidates.

Kefir, which originates in the Caucasian mountains, is a cultured milk beverage produced by a combination of acidic and alcoholic fermentation. Kefir products are commonly used as food vehicles to deliver health-promoting materials including kefran and lactic acid bacteria to consumers. The aim of this study was to develop a freeze-dried starter culture without yeast and assess the suitability of kefir-like dairy products for the growth of lactic acid bacteria and the acidification of milk. Pasteurized whole milk (SNF 8.5%) stored at 25℃ was aseptically inoculated with starter cultures (0.002% w/v); it was kept at 25℃ until the pH attained a value of 4.6. Ten grams of the kefir-like product sample was diluted with 90 mL of 0.15% peptone water diluent in a milk dilution bottle, followed by uniform mixing for 1 min. Viable cells of Lactobacillus species were enumerated on modified-MRS agar (pH 5.2), with incubation at 37℃ for 48 h. Viable cells of Lactococcus species were enumerated on M17-lactose agar, with incubation at 32℃ for 48 h. The pH attained a value of 4.6 after fermentation for 9 h 30 min (Starter 1), 9 h 45 min (Starter 2), and 12 h (Starter 3). The viable cell count of Lactobacillus sp. and Lactococcus sp. was initially 10 5 ∼10 6 CFU/g; it increased significantly to 10 9 CFU/g after 12 h of incubation. During the storage of the kefir-like products at 4℃ for 1 4 days, the total viable cell numbers were unchanged, but the pH decreased slightly. The consistency of the kefir products increased gradually during the storage. The organoleptic properties of the kefir products fermented using the new starter culture are more desirable than those of commercial kefir. These results suggest that the newly developed starter culture without yeast could be suitable for kefir fermentation.