Relationship of Cellular Fatty Acid Composition to Survival of Lactobacillus bulgaricus in Liquid Nitrogen1

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Concentrated cultures of Lactobacillus bulgaricus were prepared by resuspending cells grown in semisynthetic media in sterile 10% non-fat milk solids. The concentrated cultures were frozen in liquid nitrogen for 24 h. The cell suspensions exhibited decreased viability after storage, and the amount of death varied among the different strains tested. Storage stability of all strains examined was improved by supplementing the growth medium with sodium oleate. Radioisotopes were used to study the fate of sodium oleate with L. bulgaricus NCS1. [1-14C]sodium oleate was incorporated solely into the lipid portion of the cells, including both neutral and polar lipids. The fatty acid composition of L. bulgaricus NCS1, NCS2, NCS3, and NCS4 grown with and without sodium oleate was studied. The major fatty acids of strains NCS1, NCS2, and NCS3 grown without sodium oleate were dodecanoic, tetradecanoic, hexadecanoic, hexadecenoic, and octadecenoic acids. In addition to these, strain NCS4 contained C19 cyclopropane fatty acid. The major fatty acids of all strains grown with sodium oleate were tetradecanoic, hexadecanoic, hexadecenoic, octadecenoic, and C19 cyclopropane fatty acids. All strains grown in broth containing sodium oleate contained larger amounts of octadecenoic and C19 cyclopropane fatty acid, and less saturated fatty acids than when grown without sodium oleate. Statistical analyses indicated that C19 cyclopropane fatty acid was most closely related to stability of the lactobacilli in liquid nitrogen. A negative regression line that was significant at P < 0.001 was obtained when the cellular content of this fatty acid was plotted against death.

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