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One hundred and three strains of lactic acid bacteria, isolated from various food ecosystems, were assayed for β-glucosidase activity toward p-nitrophenyl-β-D-glucopyranoside substrate. Lactobacillus plantarum DPPMA24W and DPPMASL33, Lactobacillus fermentum DPPMA114, and Lactobacillus rhamnosus DPPMAAZ1 showed the highest activities and were selected as the mixed starter to ferment various soy milk preparations, which mainly differed for chemical composition, protein dispersibility index, and size dimension. The soy milk made with organically farmed soybeans (OFS) was selected as the best preparation. All selected strains grew well in OFS soy milk, reaching almost the same values of cell density (ca. 8.5 log cfu/mL). After 96 h of fermentation with the selected mixed starter, OFS soy milk contained 57.0 μM daidzein, 140.3 μM genistein, 20.4 μM glycitein, and 37.3 μM equol. Fermented and nonfermented OFS soy milks were used for the in vitro assays on intestinal human Caco-2/TC7 cells. Fermented OFS soy milk markedly inhibited the inflammatory status of Caco-2/TC7 cells as induced by treatment with interferon-γ (IFN-γ) (1000 U/mL) and lipopolysaccharide (LPS) (100 ng/mL), maintained the integrity of the tight junctions, even if subjected to negative stimulation by IFN-γ, and markedly inhibited the synthesis of IL-8, after treatment with interleukin-1β (2 ng/mL). As shown by using chemical standards, these effects were due to the concomitant activities of isoflavone aglycones and, especially, equol, which were synthesized in the fermented OFS soy milk preparation.

A pool of selected lactic acid bacteria was used to ferment durum wheat semolina under liquid conditions. After fermentation, the dough was freeze-dried, mixed with buckwheat flour at a ratio of 3:7, and used to produce the "fusilli" type Italian pasta. Pasta without prefermentation was used as the control. Ingredients and pastas were characterized for compositional analysis. As shown by two-dimensional electrophoresis, 92 of the 130 durum wheat gliadin spots were hydrolyzed almost totally during fermentation by lactic acid bacteria. Mass spectrometry matrix-assisted laser desorption/ionization time-of-flight and reversed phase high-performance liquid chromatography analyses confirmed the hydrolysis of gliadins. As shown by immunological analysis by R5-Western blot, the concentration of gluten decreased from 6280 ppm in the control pasta to 1045 ppm in the pasta fermented with lactic acid bacteria. Gliadins were extracted from fermented and nonfermented durum wheat dough semolina and used to produce a peptic-tryptic (PT) digest for in vitro agglutination tests on cells of human origin. The whole PT digests did not cause agglutination. Affinity chromatography on Sepharose-6-B mannan column separated the PT digests in three fractions. Fraction C showed agglutination activity. The minimal agglutinating activity of fraction C from the PT digest of fermented durum wheat semolina was ca. 80 times higher than that of durum wheat semolina. Pasta was subjected to sensory analysis: The scores for stickiness and firmness were slightly lower than those found for the pasta control. Odor and flavor did not differ between the two types of pasta. These results showed that a pasta biotechnology that uses a prefermentation of durum wheat semolina by selected lactic acid bacteria and tolerated buckwheat flour could be considered as a novel tool to potentially decrease gluten intolerance and the risk of gluten contamination in gluten-free products.