Effects of polyunsaturated fatty acids in growth medium on lipid composition and on physicochemical surface properties of lactobacilli

Abstract

Most probiotic lactobacilli adhere to intestinal surfaces, a phenomenon influenced by free polyunsaturated fatty acids (PUFA). The present study investigated whether free linoleic acid, gamma-linolenic acid, arachidonic acid, alpha-linolenic acid, or docosahexaenoic acid in the growth medium alters the fatty acid composition of lactobacilli and their physical characteristics. The most abundant bacterial fatty acids identified were oleic, vaccenic, and dihydrosterculic acids. PUFA, especially conjugated linoleic acid (CLA) isomers and gamma-linolenic, eicosapentaenoic, docosahexaenoic, and alpha-linolenic acids, also were identified in lactobacilli. When lactobacilli were cultured in MRS broth supplemented with various free PUFA, the incorporation of a given PUFA into bacterial fatty acids was clearly observed. Moreover, PUFA supplementation also resulted in PUFA-dependent changes in the proportions of other fatty acids; major interconversions were seen in octadecanoic acids (18:1), their methylenated derivatives (19:cyc), and CLA. Intermittent changes in eicosapentaenoic acid proportions also were noted. These results were paralleled by minor changes in the hydrophilic or hydrophobic characteristics of lactobacilli, suggesting that PUFA interfere with microbial adhesion to intestinal surfaces through other mechanisms. In conclusion, we have demonstrated that free PUFA in the growth medium induce changes in bacterial fatty acids in relation to the regulation of the degree of fatty acid unsaturation, cyclization, and proportions of CLA and PUFA containing 20 to 22 carbons. The potential role of lactobacilli as regulators of PUFA absorption may represent another means by which probiotics could redirect the delicate balance of inflammatory mediators derived from PUFA within the inflamed intestine.

FIG. 1.

Enlarged chromatogram of the region of the 18:0, 18:1, and CLA isomers. The chromatogram represents the cellular fatty acids of Lactobacillus GG cultured in standard, non-PUFA-supplemented MRS broth. The cyclopropenic acid region of another chromatogram (Lactobacillus GG cultured in α-linolenic acid-supplemented MRS broth) is shown in the inset. An asterisk indicates identification of a peak not based on the standard.

FIG. 2.

Solvent ratios. (A) The baseline represents the chloroform (CF)/tetradecane (C₁₄) ratio for control cultures (no PUFA supplementation), and the bars represent the differences in CF/C₁₄ ratios between test cultures (different PUFA supplementations) and control cultures; i.e., values below the baseline indicate that the CF/C₁₄ ratio of control cultures was higher than that of test cultures (suggesting decreased basic characteristics of the bacterial surface) and vice versa. (B) The baseline represents the ethyl acetate (EA)/octane (C₈) ratio for control cultures, and the bars represent the differences in EA/C₈ ratios between test cultures and control cultures; i.e., values below the baseline indicate that the EA/C₈ ratio of control cultures was higher than that of test cultures (suggesting increased acidic characteristics of the bacterial surface) and vice versa. GG, Lactobacillus GG; Shi, L. casei Shirota; Lb, L. delbrueckii subsp. bulgaricus. The x axes indicate culture conditions. Data are reported as the mean and standard error of the mean. Significant differences (P < 0.05) are marked by asterisks.