Addition of disaccharides and dimethyl sulfoxide to sodium alginate gel improves viability of immobilized Saccharomyces boulardii after cryopreservation

Abstract

Gel-immobilized microorganisms are increasingly used in microbiological industries. Currently, the problem of developing technologies for long-term storage of microorganisms immobilized in gel carriers remains urgent. Low-temperature storage is the most effective method of preserving microorganisms. The viability of immobilized cells is affected by cryoprotective properties of gel matrix as well as cooling regimes. Therefore, the effects of incorporation of cryoprotective agents in alginate gel and cooling regimes on the viability of immobilized Saccharomyces boulardii cells after cryopreservation were studied. Incorporation of non-permeable cryoprotectants (sucrose, lactose, and trehalose) and permeable one dimethyl sulfoxide (DMSO) in alginate gel beads promoted an increase in the viability of immobilized cells after freeze-thawing. The highest viability rates of the gel-immobilized cells were observed in the alginate gel beads incorporating combinations of DMSO (5 – 10 % v/v) and one of the disaccharides (10 – 20 % w/v). In all experiments, slow cooling provided significantly higher viability if compared to the rapid immersion of the samples into liquid nitrogen.