Fermented milks with specific Lactobacillus spp. with potential cardioprotective effects.

In vitro and in vivo studies have reported the potential cardioprotective effects of fermented milks (FM). The aim of the present study was to evaluate the inhibitory activities of angiotensin converting enzyme (ACE), thrombin enzyme (TI) and micellar solubility of cholesterol of FM after 24 and 48 h of fermentation with Limosilactobacillus fermentum (J20, J23, J28 and J38), Lactiplantibacillus plantarum (J25) or Lactiplantibacillus pentosus (J34 and J37) exposed to simulated gastrointestinal digestion. Results showed that FM with J20 and J23 at 48 h of fermentation presented significantly (p < 0.05) higher degree of hydrolysis than other FM, and were not significantly different (p > 0.05) between them. Conversely, peptide relative abundance was significantly (p < 0.05) higher in FM with J20 than FM with J23. Moreover, IC50 (protein concentration necessary to inhibit enzyme activity by 50%) for ACE inhibition were 0.33 and 0.5 mg/mL for FM with J20 and J23, respectively. For TI inhibition, the IC50 were 0.3 and 0.24 mg/mL for FM with J20 and J23, respectively. Results exhibited 51 and 74% inhibition of micellar solubility cholesterol for FM with J20 and J23, respectively. Therefore, these results showed that not only peptide abundance, but also specific peptides might be responsible for these potential cardioprotective effects.

Novel bacteriocins produced by Lactobacillus fermentum strains with bacteriostatic effects in milk against selected indicator microorganisms.

The aim of this work was to isolate and characterize bacteriocins produced by 2 Lactobacillus fermentum strains isolated from artisanal Mexican Cocido cheese. Fractions (F ≤3 kDa) obtained from cell-free supernatants of Lb. fermentum strains J23 and J32 were further fractionated by reversed-phase HPLC on a C18 column. Antimicrobial activities of F ≤3 kDa and bacteriocin-containing fractions (BCF), obtained from fractionation of F ≤3 kDa against 4 indicator microorganisms, were determined by the disk diffusion method and growth inhibition in milk. Subsequently, isolated BCF were analyzed by reversed-phase HPLC tandem mass spectrometry. Results showed that BCF presented antimicrobial activity against the 4 indicator microorganisms tested. For J23, one of the fractions (F3) presented the highest activity against Escherichia coli and was also inhibitory against Staphylococcus aureus, Listeria innocua, Salmonella Typhimurium, and Salmonella Choleraesuis. Similarly, fractions F3 and F4 produced by J32 presented antimicrobial activity against all indicator microorganisms. Furthermore, generation time and growth rate showed that F3 from J23 presented significantly higher antimicrobial activity against the 4 indicator microorganisms (2 gram-positive and 2 gram-negative) when inoculated in milk compared with F3 from J32. Interestingly, this fraction presented a broader antimicrobial spectrum in milk than nisin (control). Reversed-phase HPLC tandem mass spectrometry analysis revealed the presence of several peptides in BCF; however, F3 from J23 that was the most active fraction of all presented only 1 bacteriocin. The chemical characterization of this bacteriocin suggested that it was a novel peptide with 10 hydrophobic AA residues in its sequence and a molecular weight of 2,056 Da. This bacteriocin and its producing strain, J23, may find application as a biopreservative against these indicator microorganisms in dairy products.

Cooperation between Lactococcus lactis NRRL B-50571 and NRRL B-50572 for Aroma Formation in Fermented Milk.

The aim of the present study was to characterize the aroma and volatile profiles of milk fermented by wild Lactococcus lactis NRRL B-50571 (FM-571) and NRRL B-50572 (FM-572) and co-fermented with both strains (co-FM). Milks fermented by these strains have been reported to have an antihypertensive effect, yet their sensory characteristics, which are of great importance for consumer acceptance of functional foods, have not been studied. In the study, volatiles were determined using solid-phase microextraction gas chromatography mass spectrometry (SPME-GC-MS) and aroma was determined by quantitative descriptive sensory analysis (QDA). Volatile compounds identified in FM-571, FM-572, and co-FM were mainly acids, alcohols, aldehydes, and ketones. FM-571 showed higher total relative volatile abundance than FM-572 or co-FM. Furthermore, the concentrations of specific amino acids (aa) were lower in FM-571 and co-FM than in FM-572. Thus, these results suggested that FM-571 or co-FM are more efficient in transforming specific aa into the corresponding volatiles than FM-572. Indeed, several alcohols and aldehydes, associated with the catabolism of these aa, were found in FM-571 and co-FM, but not in FM-572. Additionally, QDA showed that FM-571 and co-FM presented higher yeasty and cheesy aroma descriptors than FM-572. Also, total aroma intensity scores for FM-571 were higher than those for co-FM or FM-572. Thus, results suggested that the combination of these two specific wild L. lactis strains may complement amino acid catabolic routes that resulted in the enhancement or attenuation of aroma production of single strains, presenting new possibilities for the preparation of custom-made starter cultures.

In Vitro Antithrombotic and Hypocholesterolemic Activities of Milk Fermented with Specific Strains of Lactococcus lactis.

Milk fermented with specific lactic acid bacteria (LAB) was reported to be a rich source of metabolites, such as peptides with different biological activities that may have a positive effect on cardiovascular health. Thus, in this study, the antithrombotic and hypocholesterolemic activities of fermented milk with specific strains of Lactococcus lactis were investigated before and after exposure to a simulated gastrointestinal digestion (SGD) model. The inhibition of thrombin-induced fibrin polymerization (IC50 peptide concentration necessary to inhibit thrombin activity by 50%), anticoagulant activity, inhibition of micellar solubility of cholesterol and bile acid binding capacity of water soluble fractions (WSF) <3 kDa from fermented milk were evaluated. Results indicated that the WSF from fermented milk with Lc-572 showed antithrombotic (IC50 = 0.049 mg/mL) and hypocholesterolemic (55% inhibition of micellar solubility of cholesterol and 27% bile acid binding capacity) activities. Meanwhile, fermented milk with Lc-571 showed mainly antithrombotic activity (IC50 = 0.045 mg/mL). On the other hand, fermented milk with Lc-600 presented mainly hypocholesterolemic activity (31.4% inhibition of micellar solubility of and 70% bile acid binding capacity). Moreover, biological activities were not lost after simulated gastrointestinal digestion conditions. Thus, fermented milk with these specific L. lactis strains show potential for the development of functional foods.

Milk Fermented by Specific Lactobacillus Strains Regulates the Serum Levels of IL-6, TNF-α and IL-10 Cytokines in a LPS-Stimulated Murine Model.

Studies report that metabolites, such as peptides, present in fermented milk with specific lactic acid bacteria, may regulate cytokine production and exert an anti-inflammatory effect. Hence, the cytokine regulatory effect of fermented milk by specific Lactobacillus strains was evaluated in a lipopolysaccharide (LPS)-stimulated murine model. From twelve strains, three (J20, J23 and J28) were selected for their high proteolytic and acidifying capacities in milk and used for the in vivo study. Three treatments (fermented milk, FM; pasteurized fermented milk, PFM; and its 0.05) reduced pro-inflammatory cytokine (IL-6 and TNF-α) concentrations and significantly increased anti-inflammatory (IL-10) cytokine concentrations in comparison to the control; also, pro-inflammatory cytokines were reduced for animals treated with PFM10 (p < 0.05). RP-HPLC-MS/MS analysis showed that water-soluble extracts (.

Antioxidant and antimicrobial activity of commercial propolis extract in beef patties.

The objective of this study was to evaluate the efficacy of propolis extract (PE) to reduce lipid oxidation and microbial growth on beef patties during refrigerated storage. Beef patties were manufactured by incorporating PE in 4 different treatments: (1) Control (no PE addition); (2) commercial propolis 1 (2% w/w; CP1); (3) commercial propolis 2 (2% w/w; CP2); and (4) noncommercial propolis (2% w/w; NCP). Raw patties were wrapped with polyvinyl chloride and stored at 2 °C for 8 d. Total phenolic content (TPC), free-radical scavenging activity (FRS), and polyphenolic content of the PE were evaluated using high-performance liquid chromatography (HPLC). Lipid oxidation (thiobarbituric acid-reactive substances (TBARS), conjugated dienes (CnDs), metmyoglobin (MetMb%), pH variation, and color (L*, a*, b*, C*, and h*), and microbial growth (mesophilic and psychrotrophic bacteria) of patty samples were measured. NCP treatment demonstrated the highest FRS (64.8% at 100 µg/mL), which correlated with TPC and the presence of polyphenolic compounds. Lipid oxidation (78.54%, TBARS; 45.53%, CnD; 58.57%, MetMb) and microbial mesophilic and psychrotrophic growth (19.75 and 27.03%, respectively) values were reduced by NCP treatment in refrigerated samples after 8 d. These results indicate that PE has great potential as a natural antioxidant and antimicrobial additive to extend the shelf life of beef patties.

Comparison of the milk-clotting properties of three plant extracts.

Several proteases from plant sources have been proposed as milk coagulants, however, limited research has been done on their milk-clotting properties. The effect of temperature on the milk-clotting activity of kiwi fruit, melon and ginger extracts was evaluated, as well as the effects of the different extracts on curd properties. Melon extracts showed high milk-clotting activity over a broad temperature range (45-75 °C) while kiwi fruit and ginger extracts showed high activity over a narrower temperature range, with a maximum at 40 and 63 °C, respectively. Curds produced using kiwi extracts had textural properties comparable with those obtained using commercial rennet, while melon extracts produced a fragile gel and low curd yield. The milk-clotting behavior of the three plant extracts was related to the protease specificity present in these extracts. The kiwi proteases displayed chymosin-like properties and thus hold the best potential for use as a milk coagulant in cheese production.