Background: Oxidative stress and inflammation are inextricably linked and play major roles in the onset and development of Non-communicable diseases (NCD) which are the most common cause of death and disability in modern world. Hydrolyzed proteins have also been suggested to be used to manage adverse food allergic reaction. Therefore, this study aimed to investigate anti-inflammatory and anti-allergy activities of dark muscle tuna hydrolysates using biological cell line systems as a function of enzyme, the extent of hydrolysis and molecular weight range.Methods: Dark muscle tuna hydrolysates were prepared with two different enzyme types; Alcalase and Flavourzyme. Anti-inflammation activity was measured by inhibitory effect of nitric oxide (NO) production on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. Anti-allergy was determined from ability of hydrolysates to inhibit b-hexsosaminidase (b-HEX) release from RBL-2H3 mast cells. Cytotoxicity was also investigated in both RAW 264.7 macrophage cells and RBL-2H3 mast cells.Results: No cytotoxic effect on RAW 264.7 macrophage cells and RBL-2H3 mast cells was observed. The NO inhibition and b-HEX release were found significant in dose dependent manner (p<0.05). Alcalase hydrolysates demonstrated greater anti-inflammatory and anti-allergic activities than Flavourzyme hydrolysates (p<0.05). IC50 of both effects were lower than the unhydrolyzed control, > 45.44 mg/ml for NO inhibition and > 65.23 mg/ml for b-HEX release inhibition. These effects increased with the extent of hydrolysis and enzyme concentration. The peptide of lowest molecular weight range (< 3 KDa) was highest in anti-inflammatory and anti-allergic actions. Reducing secretion of TNF-a, IL-6 and IL-1b was found greater in Alcalase hydrolysate than Flavourzyme one.Conclusions: Skipjack tuna dark muscle hydrolysates from Alcalase resulted in peptides with anti-inflammation activity, as determined by NO production in LPS-stimulated RAW 264.7 macrophage cells and anti-allergic properties as measured by a suppression of degranulation of sensitized RBL-2H3 cells. Anti-inflammatory effect may be due to their anti-oxidative capacity and relevant inflammatory factors attenuated with hydrolysate by reducing secretion of pro-inflammatory cytokine (TNF-a, IL-6 and IL-1b). Inhibition of b-HEX release by peptides may be due to membrane-stabilizing action or/and blockade of IgE antibody at fragment region.Keywords: Skipjack tuna, anti-inflammation, enzymatic hydrolysate, dark muscle, anti-allergy
Background: Royal jelly (RJ) is one of the most effectual and beneficial remedies for human beings and currently utilized in many sectors, ranging from the pharmaceutical and food industries to cosmetic and manufacturing sectors due to RJ possessing many bio-therapeutical activities including anti-tumor, antimicrobial and antioxidant activities, vasodilative and hypotensive activities, as well as growth-stimulating, infection-preventing, anti-hypercholesterolemic and anti-inflammatory activities. However, some reports showing direct consumption of RJ can lead to severe allergic reaction and has been linked with acute asthma, dermatitis, and life-threatening anaphylaxis. Thus, this research purposes to explore the potential anti-inflammatory and anti-allergic activities of hydrolyzed RJ as a function of enzyme and the extent of hydrolysis.Methods: RJ was enzymatically hydrolyzed with three commercial enzymes (AlcalaseÒ, FlavourzymeÒ and ProtamexÒ). Anti-inflammatory activity of the hydrolysates was measured by their inhibitory effect on nitric oxide (NO) production of lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Anti-allergy was determined from the ability of the hydrolysates to inhibit b-hexsosaminidase (b-HEX) release from RBL-2H3 mast cells. Cytotoxicity was also investigated in both RAW264.7 macrophage cells and RBL-2H3 mast cells.Results: The electrophoretic profiles indicated that AlcalaseÒ and FlavourzymeÒ hydrolysates did not show the presence of proteins causing allergic reaction after 60 mins of hydrolysis while these allergens disappeared from ProtamexÒ hydrolysate at the hydrolysis time of 240 min. It was observed that hydrolyzed RJ showed no toxicity on RAW264.7 and RBL-2H3 cells. With the progression of hydrolysis, IC50 values of NO production inhibition significantly decreased while degree of hydrolysis (DH) was increased in all hydrolyzed samples (p < 0.05). Results of b-HEX release inhibition were found in the same fashion. FlavourzymeÒ hydrolysate at the 240 min time point effectively mitigated the oxidative stress and protected DNA in a dose dependent manner.Conclusions: RJ hydrolysates from FlavourzymeÒ resulted in peptides with anti-inflammatory activity as determined by the inhibition of NO production in LPS-stimulated RAW264.7 macrophage cells and anti-allergic property as measured by the suppression of degranulation of sensitized RBL-2H3 cells. Anti-inflammatory effect may be due to their anti-oxidative capability. Inhibition of b-HEX release may be due to their membrane-stabilizing effects or/and blockade of IgE antibody binding to its receptors.Keywords: anti-inflammation, enzymatic hydrolysate, royal jelly, anti-allergy
Recently, consumer awareness of healthy foods and eating habits has generated a massive market demand for functional foods with health benefits. Fermented foods, particularly non-dairy beverages, are gaining popularity and acceptance due to their functional benefits [1].Biotransformation based on fermentation is controlled by two main factors, microorganisms and substrates [2]. Lactic acid bacteria (LAB) and yeast are the most used probiotics in fermented foods. Some species of yeast, including Saccharomyces cerevisiae var. boulardii (Saccharomyces boulardii), and LAB, such as L. plantarum and L. salivarius, showed positive probiotic results in the inhibition of pathogenic bacteria growth and antibiotic resistance [3]. To improve the nutritional value and organoleptic characteristics of fruit fermented juice, mixing of yeast and LAB as a multi-strain starter may provide better beneficial effects than mono-strain culture [4]. During the fermentation process, both yeast and LAB are the two main microbial groups involved in the ester and terpene production. Yeast is responsible for alcoholic fermentation and determines the production of major alcohol and esters [5]. The components of alcoholic beverages can be split into major and minor groups. The major group consists of ethanol and water, and the minor group comprises fusel alcohols, carbonyl compounds, esters, organic acids, aldehydes, lactones, and sulfur compounds [6]. Through biotransformation, LAB can produce the volatile flavors of fermented foods, which are composed of organic acids, alcohols, and ketone and aldehyde compounds [7]. When LAB and yeast were cocultured, they enhanced the growth of both groups as well as the flavor, thereby Fermentation is an effective process for providing various beneficial effects in functional beverages. Lactic acid bacteria and yeast fermentation-based biotransformation contribute to enhancement of nutritional value and digestibility, including lactose intolerance reduction and control of infections.In this study, the probiotic fermented fruit juice (PFJ) was produced by Lactobacillus plantarum TISTR 1465, Lactobacillus salivarius TISTR 1112, and Saccharomyces boulardii CNCM I-745 while mixed fruit juice (MFJ) was used as the basic medium for microorganism growth. The potential function, the anti-salmonella activity of PFJ, was found to be effective at 250 mg/ml of MIC and 500 mg/ml of MBC. Biofilm inhibition was performed using the PFJ samples and showed at least 70% reduction in cell attachment at the MIC concentration of Salmonella Typhi DMST 22842. The antioxidant activities of PFJ were determined and the results revealed that FSB.25 exhibited 78.40 ± 0.51 mM TE/ml by FRAP assay, while FPSB.25 exhibited 3.44 ± 0.10 mM TE/ml by DPPH assay. The volatile compounds of PFJ were characterized by GC-MS, which identified alcohol, aldehyde, acid, ester, ketone, phenol, and terpene. The most abundant organic acid and alcohol detected in PFJ were acetic acid and 2phenylethanol, and the most represented terpene was β-damascenone. ...
Background: Lactic acid bacteria-based fermentation clearly contributes to improving nutritional value and exhibits various health benefits. The demand for non-dairy functional beverages, such as fruit beverages, as an alternative vehicle for probiotics is increasing because of lifestyle choices or health conditions. Therefore, the objective of this study was to evaluate the anti-Salmonella potential and antioxidant activity of fermented fruit-based juice by lactic acid bacteria and its biotransformation. Methods: In this study, to produce the fermented fruit-based juice (FFJ), the mixed fruit juice (MFJ) was fermented by Lactobacillus plantarum TISTR 1465 and Lactobacillus salivarius TIST 1112 for 72 hrs. The potential function, anti-Salmonella by the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and antibiofilm activities of FFJ against Salmonella Typhi DMST 22842 was evaluated. The antioxidative capacity was determined by DPPH and FRAP assay. The active volatile compounds were identified by GC-MS. Results: A novel functional FFJ showed excellent growth capacity with 8 log CFU/mL of probiotics Lactobacillus plantarum TISTR 1465 and Lactobacillus salivarius TIST 1112. MIC and MBC values in the FFJ were 500 mg/mL after 72 hrs of fermentation. After 48hrs of fermentation, biofilm formation inhibition was significant (p < 0.05) with 95.27% ± 2.26% inhibition; biofilm metabolic activity inhibition was also significant (p < 0.05) with 89.25% ± 0.18% inhibition. The volatile compounds present in the FFJ were fruity flavors and aromas, most of have antimicrobial and antioxidant properties. These compounds comprise various classes, including alcohols, organic acid, ester, and ketone. In both LAB fermentations, the most abundant volatile alcohol was isoamyl alcohol, followed by 1-hexanol and 2,3-Butanolone; acetic acid was only present in L. plantarum fermentation. In addition, DPPH radical scavenging and FRAP assay showed the mixed fruit juice had dramatically increased antioxidant activity after 48 hrs of fermentation.Conclusion: The findings of this work indicate that the obtained fermented fruit-based juice (FFJ) showed excellent growth capacity of probiotics, Lactobacillus plantarum TISTR 1465 and Lactobacillus salivarius TIST 1112, and produced the volatile compounds from biotransformation. This not only improved fruit flavor and aroma, but also influenced antibacterial activity against the pathogen Salmonella Typhi DMST 22842, as well as increased antioxidant activity. Therefore, the FFJ could be a novel functional fermented drink for vegan and non-diary consumption.Keywords: Lactic acid bacteria, Probiotics, Biotransformation, Non-dairy functional beverage, Anti-Salmonella
Background: Potent calcium uptake is essential for calcium balance and normal health. Prolonged low intake of calcium is associated with osteoporosis, dental changes, cataracts, and alterations in the brain. However, calcium is difficult to be directly absorbed from the food due to the insoluble calcium salt precipitation that occurs in the intestinal environment. Methods: Tilapia protein hydrolysate (TPH) was prepared by alcalase digestion. The Calcium-binding activity was measured using calcium colorimetric assay, the absorption at 612 nm. The interaction between TPH and calcium was examined by spectroscopic analysis, ultraviolet absorption and fluorescence measurement. TPH-calcium-binding stability in the human digestion system was evaluated by in vitro pepsin-pancreatin hydrolysis simulating human gastric and intestinal digestion. The effects of food components on TPH-calcium-binding activity was also analyzed. The enhancement of transepithelial calcium transport by TPH was determined by in vitro Caco2 epithelial cell-like monolayer. Results: TPH produced from Nile tilapia (Oreochromis niloticus) exhibited calcium-binding activity. It was the peptides in the hydrolysate that contributed to calcium-binding since the spectroscopic changes induced by calcium were characteristic of peptide bonds and tryptophan residues. The calcium binding of TPH was compatible with food matrices. Most food components including saccharides, amino acids and vitamins showed positive or no effects on calcium-binding. The calcium-binding of TPH was also stable in the simulated gastrointestinal digestion system. Pepsin and pancreatin did not considerably change the calcium-binding activity of TPH. Of note, TPH reduced precipitation of calcium by oxalate and phytate, the two most anti-nutritional factors present in green leafy vegetables. Finally, we showed that TPH significantly promoted transepithelial calcium transport in the Caco-2 cell permeability model. Conclusions: Tilapia protein hydrolysate produced by alcalase digestion possessed calcium-binding activity and prevent precipitation of calcium by a mineral chelating agent as well as enhanced transepithelial calcium transport in Caco2 cell. The result implicated the potential of TPH as a functional food ingredient for promoting calcium absorption. Keywords: Tilapia protein hydrolysate; Calcium binding peptides; Calcium absorption
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