Effect of Low-Temperature Storage of Kefir Grains and Trehalose Addition on the Production of the Exopolysaccharide Kefiran

Kefiran, the extracellular polysaccharide produced by Generally Recognized as Safe (GRAS) bacteria found in kefir grains, is a promising biopolymer with multiple applications in agri-food and biomedical fields. Besides its characteristics and potential applications, the factors that affect its production remain a prime subject of interest. Lactic acid bacteria synthesize polysaccharides to protect their cells from adverse conditions. Therefore, low-temperature storage (4 °C) of kefir grains inoculated into Ultra-High-Temperature (UHT) milk at two different concentrations (5% and 30%) was studied as a factor for increasing kefiran production in the medium. The cryoprotectant disaccharide trehalose, which comprises a carbon and energy source for many microorganisms, was also evaluated for its effectiveness in enhancing kefiran production. The pH, the increase in kefir grain mass, the amount of kefiran produced, and the rheological properties of the acidified milk were determined during two distinct storage periods, depending on kefir grain concentration. For comparison, kefir grains were also fermented at 25 °C and 30 °C. Low-temperature storage at a kefir grain concentration of 30% resulted in an increase in the amount of polysaccharide produced beyond that obtained through fermentation. Fermentation of a 5% grain inoculum at 30 °C resulted in the lowest kefiran production. In the presence of trehalose, prolonged low-temperature storage favored an increase in the biosynthesis of kefiran, especially at a 30% kefir grain inoculum. Trehalose, however, was not a significant factor in the fermentation experiments. Proper selection of low-temperature storage time is required to avoid a reduction in kefiran concentration due to the metabolic activity of the microorganisms in kefir grains. The acidified milk (low-temperature storage) and kefir (fermentation) samples both exhibited increased elasticity and apparent viscosity with increasing kefir grain concentration. However, the rheological behavior of acidified milk was greatly affected by protein degradation during low-temperature storage. As shown by the findings of the present study, low-temperature storage (4 °C) of a 30% kefir grain inoculum in the presence of trehalose (3% w/w) until a final pH of 4.2 proves to favor kefiran production in the medium the most.