Profile of Bioactive Components of Cocoa (Theobroma cacao L.) By-Products from Ecuador and Evaluation of Their Antioxidant Activity.

The aim of the study was to determine the profile of bioactive compounds in cocoa residues (mucilage and bean shells), and to evaluate their antioxidant activity in two cocoa varieties, Nacional X Trinitario type (Fine Aroma) and the variety CCN-51. The extraction of phytonutrients from the residues was carried out selectively. The characterization and quantification of the total polyphenol content (TPC), and the total flavonoid content (TFC) were determined by UV-VIS spectrophotometry. High-performance liquid chromatography (HPLC) was used to determine the phenolic profile and methylxanthines. The antioxidant activity was evaluated by the methods of 2-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid) cation bleaching (ABTS), ferric-reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC). The exudate mucilage samples from Nacional X Trinitario-type cocoa presented the highest content of TPC 105.08 mg gallic acid equivalents (GAE)/100 mL, TFC 36.80 mg catechin equivalents (CE)/100 mL, catechin (CAT) 35.44 mg/g, procyanidins (PCB2: 35.10; PCB1: 25.68; PCC1: 16.83 mg/L), epicatechin (EPI) 13.71 mg/L, caffeine (CAF) 0.90% and theobromine (TBR) 2.65%. In the cocoa bean shell, the variety CCN-51 presented a higher content of TPC (42.17 mg GAE/100 g) and TFC (20.57 mg CE/100 g). However, CAT (16.16 mg/g), CAF (0.35%) and TBR (1.28%) were higher in the Nacional X Trinitario cocoa type. The EPI presented no significant differences between the two samples studied (0.83 and 0.84 mg/g). The antioxidant activity values (ABTS, FRAP and ORAC methods) were higher in the samples of CCN-51 than in the Nacional X Trinitario type. The bean shell samples presented antioxidant values of 171.32, 192.22 and 56.87 mg Trolox equivalents (TE)/g, respectively, and the bean shell samples presented antioxidant values of 167.06, 160.06 and 52.53 mg TE/g, respectively. The antioxidant activity (ABTS, FRAP and ORAC) of the residues was correlated with the bioactive compounds of the mucilage and bean shells, showing a strong positive correlation (<0.99) with the procyanidins (B1, B2 and C1), EPI and CAT and a positive/moderate correlation (0.94) with methylxanthines.

Polyphenols and Flavonoids Composition, Anti-Inflammatory and Antioxidant Properties of Andean Baccharis macrantha Extracts.

This study examined the leaves of Baccharis macrantha to obtain extracts of Baccharis macrantha (EBM) and to determine the total flavonoid content (TFC) and the total polyphenol content (TPC). The main objective of this work was to quantify TPC and TFC of extracts of B. macrantha from Ecuador and evaluate its antioxidant and anti-inflammatory activities and inhibition of lipid peroxidation. The extraction method was optimized with solvents, ethanol, and methanol, at temperatures of 30-60 °C and extraction times of 5-20 min. The optimal TFC extraction conditions were at EtOH25% at 50 °C for 10 min. The optimal TPC extraction conditions were at EtOH50% at 50 °C for 10 min. EBM was characterized by TLC and HPLC with three standards: gallic acid, catechin, and quercetin. EBM-EtOH25% and EBM-EtOH50% obtained at 50 °C for 10 min were used to identify quercetin and evaluate biologicals activities. Quercetin was detected in EBM (EtOH25% and EtOH50%). EBM anti-inflammatory activity was evaluated with the red blood cell stabilization (RBC) method. The RBC model showed values of 49.72% of protection lysis RBC to EBM-EtOH25% and 50.71% of protection lysis RBC to EBM-EtOH50%. The EBM in vitro inhibition of lipid peroxidation showed a protection of 77.00% (EtOH25%) and 73.11% (EtOH50%) when the TBARs method was used. EBM-EtOH25% and EtOH50% showed high antioxidant activity. EBM-EtOH25% presented values of ABTS (1172 µmol TE/g EBM), DPPH (836 µmol TE/g, EBM), and FRAP (85.70 µmol TE/g, EBM).

Functional, Antioxidant, and Anti-Inflammatory Properties of Cricket Protein Concentrate (Gryllus assimilis).

Edible insects can represent an alternative to obtain high-quality proteins with positive biological properties for human consumption. Cricket flour (Gryllus assimilis) was used to obtain cricket protein concentrate (CPC) using pHs (10.0 and 12.0) of extraction and pHs (3.0, 4.0, 5.0, and 6.0) of isoelectric precipitation (pI). Protein content, water and oil absorption capacity, protein solubility, antioxidant, and anti-inflammatory activities were determined. In addition, the protein profile was characterized by electrophoresis and the in vitro CPC digestibility was evaluated. Cricket flour presented 45.75% of protein content and CPC 12-5.0 presented a value of 71.16% protein content using the Dumas method. All samples were more soluble at pH 9.0 and 12.0. CPC 12-3.0 presented a percentage of water-binding capacity (WBC) of 41.25%. CPC 12-6.0 presented a percentage of oil-binding capacity (OBC) of 72.93%. All samples presented a high antioxidant and anti-inflammatory activity. CPC 12-4.0 presented a value FRAP of 70,034 umol trolox equivalents (TE)/g CPC, CPC 12-6.0 presented a value ABTS of 124,300 umol TE/g CPC and CPC 10-3.0 presented a DPPH value of 68,009 umol TE/g CPC. CPC 10-6.0 and CPC 12-6.0 presented high anti-inflammatory activity, with values of 93.55% and 93.15% of protection, respectively. CPCs can be used as functional ingredients in the food industry for their excellent functional and biological properties.

Effect of the growing area on the fat content and the fatty acid composition of Ecuadorian cocoa beans.

Cocoa presents a high fat content and a unique fatty acid profile defining its technological and nutritional properties. This study evaluated the fat content and fatty acid composition of Nacional cocoas, a worldwide recognised "fine" variety, collected in 85 Ecuadorian farms while taking into account 3 geographical levels (region, province, and canton). The total fat content varied from 45.61 ± 1.27 to 52.13 ± 0.58 g/100 g DW and was higher in the provinces and cantons from the Amazonian region than in those from the Pacific Coast region. A remarkable effect of the region and the province was shown on the content of individual fatty acids of Nacional cocoa beans. Total amounts of saturated and unsaturated fatty acids also depended on the growing area. Multivariate analysis provided a comprehensive assessment of the cocoa fat composition according to the origin, which may be useful for the selection of cocoas with specific technological and nutritional characteristics.

Influence of the Maturity Stage on the Phytochemical Composition and the Antioxidant Activity of Four Andean Blackberry Cultivars (Rubus glaucus Benth) from Ecuador.

Andean blackberries (Rubus glaucus Benth) are fruits rich in phytocomponents with high antioxidant activity. In this work, the changes in the total polyphenol content (TPC), the total flavonoid content (TFC), and the total anthocyanin content (TAC) of four blackberry varieties at three maturity stages (E1-25%, E2-50%, and E3-100%) were measured. The antioxidant activity (AA) was evaluated using the 2,2'azinobis-(3-ethylbenzthiazolin 6-sulphonic acid (ABTS) and ferric reducing antioxidant power (FRAP) methods. TPC and TFC content decreased with the increase in the maturity stage. The blackberry Brazos cultivar presented TPC values of 51.26, 38.16, and 31.59 mg of gallic acid equivalents (GAE)/g dry weight (DW) at E1, E2, and E3, respectively. The TAC and soluble solids increased with the increase in the maturity stage of the fruits. The Andimora variety at E3 presented a high TPC content, and the Colombiana variety presented a high TFC content. The blackberry Colombiana cultivar presented TAC values of 1.40, 2.95, and 12.26 mg cy-3-glu/100g DW at E1, E2, and E3, respectively. The blackberry Colombiana cultivar presented a high AA value at 1278.63 µmol TE/g DW according to the ABTS method and 1284.55 µmol TE/g DW according to the FRAP method. The TPC and TFC showed a high correlation with the AA according to the ABTS and the FRAP methods. The Pearson correlation between the TFC and AA/ABTS has a value of r = 0.92. The TFC and AA/FRAP present a value of r = 0.94.

Analysis of Environmental Conditions Effect in the Phytochemical Composition of Potato (Solanum tuberosum) Cultivars.

Crop productivity and food quality are affected by environmental conditions. The objective of this work was to evaluate the effect of the environment on the concentration of phytochemical components in several potato (Solanum tuberosum) cultivars. The content of vitamin C (ascorbic acid, AA), the total carotenoids content (TCC), the total polyphenols content (TPC), and the total anthocyanins content (TAC) of 11 potatoes varieties grown in Ecuador (Cutuglahua, Pujilí, and Pilahuín) was measured by the spectrophotometric method. The antioxidant capacity (AC) of potato cultivars was evaluated by the ABTS method. The AA concentration ranged between 12.67 to 39.49 mg/100g fresh weight (FW), the TCC ranged between 50.00 and 1043.50 µg/100g FW, the TPC ranged between 0.41 and 3.25 g of gallic acid equivalents (GAE)/kg dry weight (DW), the TAC ranged between 2.74 and 172.53 µg/g FW and finally the AC ranged between 36.80 and 789.19 µg of trolox equivalents (TE)/g FW. Genotypes (G), location (L), and interaction (G x L) were significant at p < 0.01. The genotype (G) showed a greater variation in the phytochemical contents. AA and TPC showed the highest correlation with the AC. A selection of genotypes with these characteristics can be used to develop germplasms with a high AC.

Argentine Patagonia barberry chemical composition and evaluation of its antioxidant capacity.

An important portion of vitamins, minerals and polyphenols components in human diet are captured from fruit consumption. Argentinean Patagonia Berberis microphylla was characterized with the phenolic content, the proximate composition and the identification and quantification of anthocyanins, not-anthocyanins and proteins. The antioxidant capacity of berberis ethanolic extracts (EB) was determined by the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. EB was used to reduce production of reactive substances species (ROS) in zebrafish. EB presented a total polyphenols content of 1,035.03 mg GAE/100 g fresh weight (FW). EB presented an ABTS value of 116.25 ± 17 µmol TE/g FW. EB presented a DPPH value of 137.80 ± 1.90 µmol TE/g FW. EB was able of reducing the ROS in zebrafish. Berberies Protein Isolate (BPI) presented proteins with bands from 15 to 62 kDa. BPI presented an ABTS value of 593.11 ± 8.60 µmol TE/g. The BPI duodenal digest presented a value of 641.07 ± 12.60 µmol TE/g digests. PRACTICAL APPLICATIONS: The practical applications of the present study are to increase scientific knowledge for consumers about the quality and benefits of the consumption of the native fruit (Berberis microphylla) from the Patagonia region of Argentine. This work describes the protein profile of berberies, their digestibility and their antioxidant activity. This study allows to better understand the phytonutrients that make up this fruit. Future studies may identify the peptides present in hydrolyzates. The bio-compounds of this fruit could be used as functional ingredients by the food industry for different purposes.

Production of White, Red and Black Quinoa (Chenopodium quinoa Willd Var. Real) Protein Isolates and Its Hydrolysates in Germinated and Non-Germinated Quinoa Samples and Antioxidant Activity Evaluation.

Red, black and white seeds quinoa were germinated at 28 °C during 24 (G1), 48 and 72 h (G3). Red quinoa presented a higher percentage of germination with a value of 46% of germination at 72 h. Quinoa protein isolate (QPI) was obtained by alkaline extraction (pH 8.0) followed by an isoelectric precipitation (pH 4.5) from white, red and black quinoa seeds, germinated QPI-G1 or QPI-G3 and non-germinated QPI-NG, Chenopodium quinoa Willd var. Real. QPI-G1, QPI-G3 and QPI-NG were subject to a simulated gastric digestion (DG) and in vitro duodenal digestion (DD). The antioxidant activity was evaluated using the 1, 1-diphenyl-2-picryl hydrazyl (DPPH), azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and oxygen radical absorbance capacity (ORAC) methods. Gastric and duodenal digest of QPI-NG and QPI-G1 and QPI-G3 from white, red and black quinoa presented antioxidant activity. QPI-G1-DD of white quinoa presented the highest antioxidant activity with a DPPH value of 167.98 µmoL TE/g of digest, QPI-G1-DD of red quinoa with an ABTS value of 204.86 µmoL TE/g of digest and QPI-G1-DD of black quinoa with an ORAC value of 401.42 µmoL TE/g of digest. QPI-G3-DD of white quinoa presented higher antioxidant activity with a DPPH value of 186.28 µmoL TE/g of sample, QPI-G3-DD of red quinoa with an ABTS value of 144.06 µmoL TE/g of digest and QPI-G3-DD of black quinoa with an ORAC value of 395.14 µmoL TE/g of digest. The inhibition of reactive oxygen species (ROS) production in the zebrafish embryo model (Danio rerio) was evaluated. Protein profiles of QPI from white, red and black from germinated quinoa and non-germinated quinoa were similar with proteins between 10 kDa to 100 kDa with the presence of globulins 11S and 7S and 2S albumins.

Biocompounds Content Prediction in Ecuadorian Fruits Using a Mathematical Model.

Anthocyanins, carotenoids and polyphenols are biomolecules that give the characteristic color to fruits. Carotenoids relate to yellow, orange and red colors whereas anthocyanins and polyphenols mainly relate to purple and red colors. Presently, standard determination of antioxidants is carried out using relatively complex methods and techniques. The aim of this study was to develop a mathematical prediction model to relate the internal color parameters of the Amazonic fruits araza (Eugenia stipitata Mc Vaugh), Andean fruit blackberry (Rubus glaucus Benth), Andean blueberry (Vaccinium floribundum Kunth), goldenberry (Physalis peruviana L.), naranjilla (Solanum quitoense Lam.), and tamarillo (Solanum betaceum Cav.) to their respective anthocyanins, carotenoids and polyphenols contents. The mathematical model was effective in predicting the total anthocyanins content (TAC), the total carotenoids content (TCC) and finally the total phenolic content (TPC) of fruits assayed. Andean blueberry presented a TPC with an experimental value of 7254.62 (mg GAE/100 g sample) with respect to a TPC prediction value of 7315.73 (mg GAE/100 g sample). Andean blackberry presented a TAC with an experimental value of 1416.69 (mg chloride cyanidin 3-glucoside/100 g) with respect to a prediction TAC value of 1413 (mg chloride cyanidin 3-glucoside/100 g).

Antibacterial Activity of Hen Egg White Lysozyme Denatured by Thermal and Chemical Treatments.

The aim of this study was to increase the antibacterial spectrum of modified hen egg white lysozyme (HEWL) with thermal and chemical treatments against Gram-negative bacteria. The antibacterial activity of heat-denatured HEWL and chemical denatured HEWL against Gram-negative and Gram-positive bacteria was evaluated in 15 h of incubation tests. HEWL was denatured by heating at pH 6.0 and pH 7.0 and chemical denaturing was carried out for 1.0, 1.5, 2.0, and 4.0 h with DL-Dithiothreitol (DTT). HEWL modified by thermal and chemical treatments was characterized using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis method. Heat-denatured HEWL lytic activity against Micrococcus lysodeikticus lessened with increasing temperature and time of incubation with the chemical agent (DTT). The loss of lytic activity in modified HEWL suggests that the mechanism of action of the antibacterial activity is not dependent on the lytic activity. Thermal and chemical treatments of HEWL enabled the production of oligoforms and increased antibacterial activity over a wider spectrum. Heat-denatured HEWL at pH 6.0 and chemically-denatured HEWL increased the HEWL antibacterial spectrum against Gram-negative bacteria (Escherichia coli ATCC 25922). HEWL at 120 °C and pH 6.0 (1.0 mg/mL) inhibited 78.20% of the growth of E. coli. HEWL/DTT treatment for 4.0 h (1.0 mg/mL) inhibited 68.75% of the growth E. coli. Heat-denatured HEWL at pH 6.0 and pH 7.0 and chemically-denatured HEWL (1.0, 1.5, 2.0, and 4.0 h with DTT) were active against Gram-positive bacteria (Staphylococcus carnosus CECT 4491T). Heat-denatured and chemical-denatured HEWL caused the death of the bacteria with the destruction of the cell wall. LIVE/DEAD assays of fluorescent dye stain of the membrane cell showed membrane perturbation of bacteria after incubation with modified HEWL. The cell wall destruction was viewed using electron microscopy. The results obtained in this study suggest that heat-denatured HEWL at pH 6.0 and chemical-denatured HEWL treatments increase the HEWL antibacterial activity against Gram-negative bacteria.

Inhibition of Lipid Peroxidation of Kiwicha (Amaranthus caudatus) Hydrolyzed Protein Using Zebrafish Larvae and Embryos.

Amaranth protein concentrate (APC) was hydrolyzed under in vitro gastrointestinal conditions. APC proteins were partially degraded by pepsin at pHs 1.2, 2.0, and 3.2. During the intestinal phase (pepsin/pancreatin enzymes at pH 7.0), no polypeptide bands were observed in the gel, suggesting the susceptibility of amaranth proteins to the action of digestive enzymes. The potent in vitro inhibition of lipid peroxidation, shown by the gastric and intestinal digests, was confirmed in the zebrafish larvae, with a 72.86% reduction in oxidation of lipids in the presence of the gastric hydrolysate at pH 2.0, compared to a 95.72% reduction in the presence of the gastrointestinal digest. APC digests were capable of reducing reactive oxygen species (ROS) production in the zebrafish embryo model with a value of fluorescence of 52.5% for the gastric hydrolysate, and 48.4% for the intestinal hydrolysate.

Identification of Antimicrobial Peptides of Native and Heated Hydrolysates from Hen Egg White Lysozyme.

Hen eggs are a source of bioactive compounds, of which the hen egg white lysozyme (HEWL) protein. HEWL has a demonstrated antibacterial activity. The aim of this study was to evaluate the antimicrobial activity of native and heated HEWL hydrolysates obtained through hydrolysis with pepsin and to identify their peptides using the reversed phase high performance liquid chromatography-electrospray ionization-tandem mass spectrometry (RP-HPLC-ESI-MS-MS) analysis. Native and heat-treated HEWL was hydrolyzed with pepsin at pH 1.2, and their antibacterial activity was tested against Escherichia coli and Staphylococcus carnosus. Two of the hydrolysates obtained presented high antibacterial activity against Gram-positive and Gram-negative bacteria. Native HEWL hydrolysate was a bactericide at 2.0 mg/mL against E. coli. Fifty-one peptide sequences were identified on the two hydrolysates. Peptides identified are cationic peptides. These peptides are rich in Lys and Arg cationic amino acids and have Trp in their sequences.

Isolation of Antibacterial Hydrolysates from Hen Egg White Lysozyme and Identification of Antibacterial Peptides.

Native and heated hen egg white lysozyme (HEWL) hydrolysates were isolated by hydrolysis with pepsin at pH 2.0 in situ in a cation exchange membrane to isolate and identify antibacterial peptides of the HEWL hydrolysates. Native and heated HEWL was partially hydrolyzed with pepsin at pH 2.0. The fractions were eluted with 5 M ammonia to identify 23 antibacterial peptides using a tandem mass spectrometry. Then, these fractions were eluted with a solution of NaCl 1 M, and seven positively charged peptides f(23-28) YSLGNW, f(122-129) AWIRGCRL, f(123-129) WIRGCRL, f(124-129) IRGCRL, f(82-96) ALLSSDITASVNCAK, f(103-129) VAWRNRCKGTDVQAWIRGCRL, and f(97-123) KIVSDGNGMNAWVAWRNRCKGT were identified using tandem mass spectrometry. Native HEWL hydrolysate presented an enzymatic activity of 23.0%, heated HEWL hydrolysate at pH 6.0 presented a residual enzymatic activity of 22.0%, and heated HEWL hydrolysate at pH 7.0 presented an enzymatic activity of 21.33%. Native and heated HEWL hydrolysate presented antibacterial activity against Escherichia coli and Staphylococcus carnosus. Native HEWL hydrolysate presented a higher enzymatic activity than heated HEWL hydrolysates.

Antiulcerative and Antinociceptive Activities of Casein and Whey Proteins.

The aim of this study was to evaluate the antiulcerative and antinociceptive activities of milk proteins using the induced gastric ulcer with ethanol rat model and the acetic acid-induced writhing mouse model. Casein (CN), (100, 300, and 1000 mg kg-1) doses presented antiulcerative activity on a dose-dependent manner with values of 30.8%, 41.4%, and 57.0% of inhibition measured using the ulcerative lesions index (ULI), respectively. Whey protein concentrate (WPC), (100, 300, and 1000 mg kg-1) doses presented antiulcerative activity on a dose-dependent manner with values of 48.9%, 65.5%, and 68.22% of ULI inhibition, respectively. CN, casein hydrolysates (CNH), WPC, and whey protein hydrolysates (WPH), (3, 10, and 30 mg kg-1) doses presented antinociceptive activity using the acetic acid-induced writhing in the mouse model. CN (30 mg kg-1) presented a value of 40% of inhibition writhing, and CNH (30 mg kg-1) presented antinociceptive activity with a value up to 46% of writhing inhibition. WPC (30 mg kg-1) presented a value of 52.50%, and WPH (30 mg kg-1) presented antinociceptive activity with a value up to 88.00% of writhing inhibition. In conclusion, CN and WPC demonstrated in vivo antiulcerative properties and represent a promising alternative to be used as protectors of the gastric mucosa. CNH and WPH demonstrated in vivo antiulcerative properties and represent a promising alternative to be used as natural analgesic.

In vitro chemopreventive properties of peptides released from quinoa (Chenopodium quinoa Willd.) protein under simulated gastrointestinal digestion.

Because of the continuous and direct interaction between the digestive tract and foods, dietary compounds represent an interesting source of chemopreventive agents for gastrointestinal health. In this study, the influence of a standardized static in vitro gastrointestinal digestion model on the release of peptides with chemopreventive potential from quinoa protein was investigated. Gastroduodenal digests and fractions collected by ultrafiltration were evaluated for their in plate oxygen radical absorbance capacity and in vitro colon cancer cell viability inhibitory activity. Highest effects were observed in the digests obtained during the intestinal phase, with fraction containing peptides <5kDa as the main responsible for the antioxidant activity and peptides >5kDa showing the greatest anti-cancer effects. Seventeen potential bioactive peptides derived from quinoa proteins have been identified. These proteins might be utilized as new ingredients in the development of functional foods or nutraceuticals with the aim of reducing oxidative stress-associated diseases, including cancer.

Antiulcerative Activity of Milk Proteins Hydrolysates.

Several studies have shown the protective effect of dairy products, especially α-lactalbumin and derived hydrolysates, against induced gastric ulcerative lesions. The mucus strengthening represents an important mechanism in the defense of gastrointestinal mucosa. Previously, a hydrolysate from casein (CNH) and a hydrolysate from whey protein concentrate rich in ß-lactoglobulin (WPH) demonstrated a stimulatory activity on mucus production in intestinal goblet cells. The aim of this work was to evaluate the possible antiulcerative activity of these two hydrolysates in an ethanol-induced ulcer model in rats. All tested samples significantly reduced the ulcerative lesions index (ULI), compared with the saline solution, using doses of 300 and 1000 mg kg-1 body weight with decreases up to 66.3% ULI. A dose-response relationship was found for both hydrolysates. The involvement of endogenous sulfhydryl (SH) groups and prostaglandins (PGs) in the antiulcerative activity was evaluated using their blockage. The antiulcerative activity of WPH showed a drastic decrease in presence of N-ethylmaleimide (from 41.4% to 9.2% ULI). However, the CNH antiulcerative properties were not significantly affected. The cytoprotective effect of WPH appears to depend on a PG-mediated mechanism. In conclusion, CNH and WPH demonstrated in vivo antiulcerative properties and represent a promising alternative as protectors of the gastric mucosa.

Native and Heated Hydrolysates of Milk Proteins and Their Capacity to Inhibit Lipid Peroxidation in the Zebrafish Larvae Model.

Casein and whey proteins with and without heat treatment were obtained of whole milk and four commercial milks in Ecuador, and were hydrolyzed. Then, their capacity to inhibit the lipid peroxidation using the TBARS method was evaluated at concentrations of 0.02, 0.04, 0.2, and, 0.4 mg/mL. Native and heated hydrolysates of milk proteins present high inhibitions of lipid peroxidation with a dose dependent effect both in vivo and in vitro tests. Casein and whey proteins obtained from whole milk were the ones with the highest anti-oxidant activity in vitro and in vivo test. Native casein hydrolysate at 0.4 mg/mL present a value of 55.55% of inhibition of lipid peroxidation and heated casein hydrolysate at 0.4 mg/mL presents a value of 58.00% of inhibition of lipid peroxidation. Native whey protein at 0.4 mg/mL present a value of 34.84% of inhibition of lipid peroxidation, and heated whey protein at 0.4 mg/mL presents a value of 40.86% of inhibition of lipid peroxidation. Native and heated casein hydrolysates were more active than native and heated whey protein hydrolysates. Heat treatments have an effect of increasing the in vitro inhibition of lipid peroxidation of hydrolysates of milk protein. Casein and whey hydrolysates were able to inhibiting lipid peroxidation in the zebrafish larvae model. Native casein hydrolysate obtained of whole milk presents 48.35% of inhibition TBARS in vivo, this activity was higher in heated casein hydrolysate obtained of whole milk with a value of 56.28% of inhibition TBARS in vivo. Native whey protein hydrolysate obtained of whole milk presents 35.30% of inhibition TBARS, and heated whey protein hydrolysate obtained of whole milk was higher, with a value of 43.60% of inhibition TBARS in vivo.

Antiproliferative Activity of Walnut (Juglans regia L.) Proteins and Walnut Protein Hydrolysates.

Proteins from Juglans regia L. were isolated. Then, proteins were hydrolyzed with different enzymes. Antiproliferative activity of proteins and of the protein hydrolysates of J. regia L. were evaluated using the sulforhodamine B method. Glutelin and prolamin proteins presented a high antiproliferative activity against cancer cells PC-3 (prostate) and K-562 (leukemia) with values of 43.9 and 84.4 µg/mL, respectively. The highest inhibitory effect observed was 50% at 0.25 µg/mL concentration in gastrointestinal digestion with pepsin and corolase pp in a dose-dependent manner against cancer cell UACC62 (melanoma). Pepsin hydrolysate showed inhibitory effects against cancer cell UACC62 (melanoma) with a concentration of 71.0 µg/mL. The effects were studied in a dose-dependent manner. The hydrolysate obtained with neutrase enzyme presented inhibitory effects against cancer cell UACC62 (melanoma) at a concentration of 25 µg/mL. Neither proteins nor protein hydrolysates presented cytotoxicity against normal cell assay VERO (epithelial).

Anti-Inflammatory and Anti-Nociceptive Activities of Native and Modified Hen Egg White Lysozyme.

Persistent inflammatory conditions can have severe pathological consequences. Although the use of nonsteroidal anti-inflammatory drugs (NSAIDs) is effective, it has side effects, particularly at the gastrointestinal level. There is then a high interest to identify natural anti-inflammatory compounds with no side effects. The anti-inflammatory and anti-nociceptive activities of hen egg lysozyme (LZ), both in its native form and modified by heat treatment, chemically or by enzymatic digestion have been tested in this study. The carrageenan-induced model in mice using native LZ or modified LZ has been applied. It was observed that LZ denatured by heat treatment at pH 6.0 presented 39.47% of inhibition of paw edema when administered at 30 mg/kg. LZ denatured with DL-dithiothreitol (DTT) presented a significant result of 42.10% inhibition of paw edema when administered at 30 mg/kg of animal weight. Modified LZ showed anti-inflammatory capacity comparable with the activity of the positive control dexamethasone. A classical model of acetic acid-induced abdominal writhing tests in mice was used to assess anti-nociceptive activity of native LZ and denatured heat treatment LZ and denatured chemical agent LZ. Finally, hydrolyzed native LZ presented 48% of inhibition of abdominal writhing in mice. Modified LZ with heat, chemical, and hydrolysis presented anti-inflammatory and anti-nociceptive activities independently of their natural enzymatic activity. These novel data point out the potential use of denatured and digested LZ as therapeutic agents and offer alternatives to the use of NSAIDs. LZ can be a natural source of anti-inflammatory and anti-nociceptive agents.

Lisozima: Actividad antibacteriana y alergenicidad / Lysozyme: Antibacterial Activity and Allergenicity

Desde que la lisozima fue descubierta por Alexander Fleming en 1922, muchos son los trabajos que se han lleva-do a cabo para describir las distintas actividades biológicas de esta proteína como son su actividad antibacteria-na, antiviral, antinflamatoria, analgésica, antitumoral y antioxidante. Su actividad antibacteriana frente a bacterias Gram-positivas es la actividad más ampliamente estudiada. Muchas investigaciones se han realizado para ampliar el espectro antibacteriano y poder atacar bacterias Gram-negativas. Se han llevado a cabo modificacio-nes térmicas, químicas, enzimáticas, mutaciones genéticas y efectos sinérgicos con otros compuestos, y se consiguió exitosamente ampliar el espectro antibacteriano, proponiendo en todos los casos que dicha actividad es independiente de su a ctividad enzimática natural. En este trabajo destacamos todas las propiedades estructura-les de la lisozima y sus principales actividades biológicas y las investigaciones que se han llevado a cabo para potenciar dichas actividades.

Since ly sozyme was discovered by Alexander Fleming in1922, many papers have been published on the different biological activities of this protein, such as its antibacterial, antiviral, anti-inflammatory, analgesic, antitumor andantioxidant properties. Its antibacterial activity againstGram-positive bacteria is the most widely studied. Much research has been done to widen the antibacterial spectrumand to attack the Gram-negative bacteria. Thermal, chemical and enzymatic modifications, as well as genetic mutations and synergistic effects, with other compounds have been made. The antibacterial spectrum was success fully widened in all cases, suggesting that this activity is independent of its natural enzymatic activity. In this paper we describe t he structural properties of lysozyme and its main biological activities, and also the research that has been carried out to maximize these activitie.

Desde que a lisozima foi descoberta por AlexandreFleming em 1922, muitos são os trabalhos que foram realizados para descrever as diferentes atividades biológicas desta proteína, como são sua atividade bacteriana, antiviral, anti-inflamatória, analgésica, antitumoral e antioxidante. Sua atividade antibacteriana frente a bactérias Gram-positivas é a atividade mais amplamente estudada. Muitas pesquisas foram realizadas para ampliar o espectroanti bacteriano e poder atacar bactérias Gram-negativas.Foram realizadas modificações térmicas, químicas,enzimáticas, mutações genéticas e efeitos sinérgicos com outros compostos, e obteve-se com sucesso ampliar o espectro antibacteriano, propondo em todos os casos que tal atividade é independente da sua atividade enzimática natural.Neste trabalho destacamos todas as propriedades estruturais da lisozima e suas principais atividades biológicas e as pesquisas que foram realizadas para potenciar tais atividades.