Angiotensin‐converting enzyme‐inhibitory and antithrombotic activities of soluble peptide extracts from buffalo and cow milk Cheddar cheeses

The aim of the study was to evaluate potential role of a water‐soluble peptide (WSP) extracts derived from buffalo and cow milk Cheddar cheeses with special reference to their antihypertension and antithrombotic activities. The WSP fractions collected at different stages of ripening were tested to assess their degree of proteolysis, their peptides were profiled by RP‐HPLC and in vitro assays for potential bioactivity were conducted. The peptide peak development was observed with slight differences in peaks number, area and height. Both angiotensin‐converting enzyme‐inhibitory and antithrombotic activities increased progressively during ripening. In comparison, the highest activities were observed in peptide extracts obtained from buffalo milk Cheddar cheese, in a dose‐dependent fashion.     

Evaluation of anti‐proliferative activity of Cheddar cheeses using colon adenocarcinoma (HCT‐116) cell line

The present study was planned to evaluate anti‐proliferative activity of water‐soluble peptides (WSPs) extracts of Cheddar cheeses made with buffalo and cow milk using a colon adenocarcinoma cell line. Cheese extracts were prepared at different stages of ripening up to six months. Anti‐proliferative activity of extracts was evaluated through cell viability assay, cell cycle arrest and apoptosis induction using colon cancer (HCT‐116) cell line. A dose‐dependent increase in activity was observed till five months of ripening. Cells population was relatively higher at G₀/G₁ phase of cell cycle. Moreover, apoptosis induction was also observed in a dose‐dependent manner.     

Survival of micro‐organisms and organic acid profile of probiotic Cheddar cheese from buffalo milk during accelerated ripening

The study aimed to assess the impact of ripening at elevated temperatures on the survival of probiotic micro‐organisms and production of organic acids in Cheddar cheese. Cheese was manufactured from buffalo milk using lactococci starters along with different probiotic bacteria (Lactobacillus acidophilus LA‐5, Bifidobacterium bifidum Bb‐11 and Bifidobacterium longum BB536) as adjunct cultures. The cheeses were ripened at 4–6 °C or 12–14 °C for 180 days and examined for composition, organic acids and microbial survival. The production of organic acids was accelerated at 12–14 °C when compared to normal ripening temperatures. The probiotic bacteria increased production of lactic and acetic acids, compared to cheese made with lactococci alone. The survival of the mesophilic starters was significantly (P < 0.05) reduced in all the cheese samples ripened at the higher temperature. However, the probiotic bacteria remained viable (>7.0 log₁₀ cfu/g) throughout the 180 days of ripening, irrespective of temperature. It was concluded that Cheddar containing additional probiotic cultures can effectively be ripened at elevated temperatures without any adverse effects.     

Anti‐inflammatory properties of high pressure‐assisted extracts of Grifola frondosa in lipopolysaccharide‐activated RAW 264.7 macrophages

The aim of this study was to determine the in vitro anti‐inflammatory properties of the shake extract (SE) and the high pressure‐assisted extract (PE) of the mycelia of Grifola frondosa in a lipopolysaccharide‐stimulated RAW 264.7 macrophage model. The content of total polysaccharides and β‐glucans of PE at 600 MPa (PE‐600) was 41.2 and 6.2 mg g^?1 dry weight, respectively, which were significantly higher than SE extracts. The results showed that treatment with 500 μg mL^?1 of PE by 600 MPa (PE‐600) did not reduce RAW 264.7 cell viability but did significantly inhibit the production of LPS‐induced NO, PGE₂ and intracellular ROS. The PE‐600 inhibited the activation of NF‐kB and then reduced the production of LPS‐induced TNF‐α, IL‐6 and IL‐1β in a dose‐dependent manner. Thus, the PE could be used as an alternative extraction method for improving the extraction efficacy of G. frondosa and serve as an alternative source of anti‐inflammatory agents.     

Anti‐inflammatory activity of rosemary extracts obtained by supercritical carbon dioxide enriched in carnosic acid and carnosol

The in vitro anti‐inflammatory activity of supercritical rosemary (Rosmarinus officinalis L.) extracts (rosemary A and B) is been reported in this study. To achieve that, THP‐1 macrophages were activated using lipopolysaccharide or human ox‐LDL and secretion and gene expression of TNF‐α, IL‐1β, IL‐6 and IL‐10 were evaluated, as well as COX‐2 gene expression. Results indicated that both rosemary extracts (A & B) exhibit high anti‐inflammatory activity although at a higher extent in case of rosemary B extract (5 μg mL^?1), representing a higher quantity of carnosic acid and carnosol than rosemary A. When comparing the activity of the extract to the standard itself, the anti‐inflammatory activity of standards of carnosic acid and carnosol was not as intense as that obtained with rosemary B. These data indicated that although carnosic acid content in the extracts is considered as the main anti‐inflammatory compound, a synergistic interaction with other compounds may play a significant role in enhancing its activity. Results provided the grounds for possible increase in the application of supercritical rosemary extracts in food formulations for mitigation or prevention of inflammatory diseases.     

Texture,flavor, and sensory quality of buffalo milk Cheddar cheese as influenced by reducing sodium salt content

The adverse health effects of dietary sodium demand the production of cheese with reduced salt content. The study was aimed to assess the effect of reducing the level of sodium chloride on the texture, flavor, and sensory qualities of Cheddar cheese. Cheddar cheese was manufactured from buffalo milk standardized at 4% fat level by adding sodium chloride at 2.5, 2.0, 1.5, 1.0, and 0.5% (wt/wt of the curd obtained). Cheese samples were ripened at 6 to 8°C for 180 d and analyzed for chemical composition after 1 wk; for texture and proteolysis after 1, 60, 120, and 180 d; and for volatile flavor compounds and sensory quality after 180 d of ripening. Decreasing the salt level significantly reduced the salt-in-moisture and pH and increased the moisture-in-nonfat-substances and water activity. Cheese hardness, toughness, and crumbliness decreased but proteolysis increased considerably on reducing the sodium content and during cheese ripening. Lowering the salt levels appreciably enhanced the concentration of volatile compounds associated with flavor but negatively affected the sensory perception. We concluded that salt level in cheese can be successfully reduced to a great extent if proteolysis and development of off-flavors resulted by the growth of starter and nonstarter bacteria can be controlled.     

Low‐fat Cheddar cheese made using microparticulated whey proteins: Effect on yield and cheese quality

Influence of different levels (0, 0.15, 0.35 or 0.50%) of microparticulated whey protein (MWP) on yield and quality of low‐fat (~7.3 g/100 g) Cheddar cheese was investigated. MWP improved cheese yield due to the water‐binding ability of denatured whey protein. MWP addition decreased meltability but improved the textural properties beneficial for shredding and slicing, by decreasing sensory firmness. The results emphasise the role of MWP as an inert filler within cheese matrix, in improving cheese yield and creating a softer texture without compromising the sensory or overall quality of cheese, even with moisture increases in 0.35 or 0.50% MWP cheeses.     

Utilisation of a cell‐free extract of lactic acid bacteria entrapped in yeast to enhance flavour development in Cheddar cheese

A lactococcal cell‐free extract (CFE) was successfully entrapped in freeze‐dried attenuated yeast. The entrapment process involved passive diffusion of enzymes from the CFE into the yeast during hydration. The entrapped CFE was subsequently added during Cheddar cheese production and its impact on a range of ripening parameters compared to added attenuated yeast or CFE alone. Statistically significant differences were evident for secondary proteolysis, sensory attributes and volatiles, which were related to enhanced enzymatic and metabolic activity from the attenuated yeast and entrapped CFE. This study highlights the potential of attenuated yeast as a vector to augment flavour development in Cheddar cheese.     

Esterase activities of indigenous lactic acid bacteria from Argentinean goats’ milk and cheeses

Esterase activities of indigenous lactic acid bacteria isolated from goats’ milk and cheese were investigated. All of the strains exhibited esterase activity on α-naphthyl derivative of fatty acids of 2–6 carbon atoms. Lactobacillus fermentum ETC1 and L. bulgaricus ETC2 showed the highest specific activity on α-naphthyl acetate; L. rhamnosus ETC14 presented the highest specific activity on α-naphthyl butyrate and caproate. All enterococci strains presented the highest specific activities on α-naphthyl propionate, butyrate and caproate, and the lowest specific activities on α-naphthyl acetate. Pedicoccus pentocaseus ETC5 only had esterase activity on α-naphthyl acetate. The electrophoretic zymogram showed for each strain an individual enzyme profile on α-naphthyl acetate and revealed the presence of more than one esterase. L. plantarum ETC11 and Enterococcus faecium ETC9 showed four and five bands of esterase activity, respectively. The strains evaluated in this work showed different esterase activities, which were species and strain specific.     

The effect of using an exopolysaccharide‐producing culture on the physicochemical properties of low‐fat and reduced‐fat Kasar cheeses

A mixed starter culture containing exopolysaccharide (EPS)‐producing strains of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus was combined with Lactobacillus helveticus LH301 and used in the manufacture of low‐fat and reduced‐fat Kasar cheeses. For comparison, low‐fat (C10) and reduced‐fat (C20) cheeses were made using EPS‐producing (EPS⁺) starter strain and EPS‐non‐producing (EPS^?) starter strain. The physicochemical properties of the cheeses were assessed in terms of chemical composition, texture, microstructure and microbial content over 90 days. Cheeses made with EPS‐producing culture (EPS10 and EPS20) had lower protein contents than control cheeses with 10% and 20% fat in dry basis (C10 and C20). Scanning electron microscopy images showed that using EPS‐producing culture resulted in a less compact protein matrix and sponge‐like structure in the cheese samples. In general, cheeses made using EPS‐producing culture had lower total viable counts. This could be related to the reduced survivability of EPS‐producing cells in the cheese matrix during ripening due to autolysis ability.     

Manufacture of dry‐ and brine‐salted soft camel cheeses for the camel dairy industry

Two experiments were conducted in a camel cheese study to (i) compare camel cheese to bovine cheese made from bovine milk standardised to simulate camel milk, and (ii) describe the technology for manufacture of dry (SCC‐D) and brine‐salted soft camel cheese (SCC‐B). Comparable cheese yield (camel: 7.4 ± 0.15, cow: 7.3 ± 0.55 kg/100 kg of milk) and levels of dry matter loss in whey were observed. Clotting time was 234 s for both cheeses which were made using thermophillic starters. Cheese yield was 9.31 ± 0.64 kg/100 kg with 425.6 ± 38.2 g/kg cheese dry matter for SCC‐D and 8.22 ± 0.90 kg/100 kg with 469 ± 73.8 g/kg dry matter for SCC‐B.     

Antioxidant activities of pepsin hydrolysates of water‐ and salt‐soluble protein extracted from pork hams

The objective of this study was to evaluate the antioxidant activities of pepsin‐digested water‐soluble protein (WSP) and salt‐soluble protein (SSP) extracted from pork ham. WSP and SSP were hydrolysed by pepsin for 2–10 h with 2‐h increments. The protein hydrolysates by pepsin were analysed by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis, and reducing power of the hydrolysates was measured. In addition, antioxidant activity of the hydrolysates was determined using linoleic acid emulsion system. Protein bands with molecular weight higher than 7 kDa in WSP and SSP were completely hydrolysed by pepsin after 2 h of digestion time. WSP hydrolysates (WSPH) and SSP hydrolysates (SSPH) had higher ferric reducing power than controls (WSP and SSP without pepsin digestion). Reducing powers of WSPH were higher than those of SSPH, regardless of digestion time. The oxidative activity of linoleic acid was predominantly inhibited by the addition of WSPH and SSPH, especially 0.5% protein hydrolysate. These results indicate that several functional peptides of pork protein digested by pepsin might have antioxidant activity, and thus they may be used as an antioxidant agent in muscle food system.     

Protein profiles and total antioxidant capacity of water‐soluble and water‐insoluble fractions of white brined goat cheese at different stages of ripening

This study deals with proteolysis and total antioxidant capacity of proteins of white brined cheese prepared from overheated goat milk and ripened for fifty days. Proteolytic changes were reflected through the relatively low level of soluble nitrogen (50 days ripened cheese had 15.32 g/100 g of water‐soluble nitrogen, 8.1 g/100 g of TCA‐soluble nitrogen and 2.69 g/100 g of PTA‐SN), intensive proteolysis of α_s2‐CN during initial 10 days of ripening (up to 50.70% of initial content) and its much slower degradation through further 40 days, slow but continual decrease of β‐CN content (up to 85.14% of residual content) and high level of proteolytic products tightly bounded into gel network. Total antioxidant capacity of water‐soluble and water‐insoluble fractions increased after cheese ripening. These findings could be useful for better understanding and control over the production of white brined goat cheese as highly valuable functional product.     

Phytochemical Analysis and Anti‐inflammatory Potential of Hyphaene thebaica L. Fruit

Metabolite profiling and biological activity are reported from organic and aqueous extracts of the fruit from the desert palm Hyphaene thebaica. Phenolics and oxylipids profiles were determined using UPLC‐PDA‐TOF (ultra performance‐photodiode array‐time of flight) high‐resolution mass spectrometry in order to obtain the molecular formula and exact mass Under optimized conditions, 17 compounds were simultaneously identified and quantified including 2 cinnamic acid derivatives, 5 flavonoids, 6 fatty acids, 2 sphingolipids, a lignan, and a stilbene. Sugars composition in the fruit was characterized and quantified by ¹H‐NMR (nuclear magnetic resonance) with sucrose detected as the major component in fruit at a level of 219 mg/g. Fruit organic extracts anti‐inflammatory potential was assessed in vitro by cyclooxygenase‐1 enzyme inhibition.     

The determination of water‐soluble vitamins and in vitro digestibility of selected Czech cheeses

Water‐soluble vitamins, in vitro organic matter digestibility (OMD) and dry matter digestibility (DMD) values were determined in various different Czech cheeses. An HPLC method for the simultaneous determination of niacin, pantothenic acid and pyridoxine in cheese was established. Different hydrolysis conditions and enzymes were used to release these vitamins from their complex form. The cheeses contained on average 2.04 mg of niacin, 3.24 mg of pantothenic acid and 2.16 mg of pyridoxine per 100 g of fresh mass. An enzymatic method was applied for the evaluation of OMD and DMD in cheeses. In vitro digestibility values ranged from 83.29% to 99.99% for organic matter (OMD) and 83.33% to 99.87% for dry matter (DMD) after two different enzymatic digestions (pepsin, pancreatin). In addition, cheeses incubated with pepsin plus pancreatin, OMD and DMD were strongly digested (99.04–99.83%, 97.73–99.74%).