Lactobacillus sanfranciscensis DSM20451 cells containing glutathione (GSH) displayed a significantly higher resistance against cold stress induced by freeze-drying, freeze-thawing, and 4°C cold treatment. Cells containing GSH were capable of maintaining the membrane structure intact when exposed to freeze-thawing. In addition, cells containing GSH showed a higher proportions of unsaturated fatty acids in cell membranes upon long-term cold treatment. Subsequent study reveals that the protective role of GSH on cell membrane against cryodamage is partly due to preventing peroxidation of membrane fatty acids and protecting Na⁺, K⁺-ATPase. The intracellular accumulation of GSH enhanced the survival and the biotechnological performance of L. sanfranciscensis, suggesting the robustness of starters for sourdough fermentation can be improved by selecting GSH accumulating strains. Moreover, this study may represent a further example of mechanisms for stress responses in lactic acid bacteria.