乳源抗氧化活性肽的研究进展

乳源抗氧化肽具有原料来源广泛、抗氧化活性强、天然无毒副作用等优点,已成为国内外学者的研究热点。为此,本文综述了乳源抗氧化肽的研究概况。首先探讨了乳源抗氧化肽的六大制备途径,发现酶解制备法使用最多。再总结了分离纯化的方法,一般为几种方法结合使用,层层递进的得到纯度较高的抗氧化肽。比较了多种抗氧化活性测定方法,在乳源抗氧化肽领域使用最多的是体外自由基清除率的测定方法,并总结了抗氧化肽分子特征,主要是由2~20个氨基酸组成的小肽,分子量在5000 Da以下,肽段中疏水性氨基酸和芳香族氨基酸占优势,但具体的构效关系还未明确。阐述了其在食品当中的应用研究现状,并展望了将来的研究及应用方向。乳源抗氧化肽显示出一定的优势,具有广阔的市场前景。     

Bioactive peptides with ACE-I and antioxidant activity produced from milk proteolysis

In this study, milk was hydrolyzed using protease (Asperigillus oryzae), trypsin, pepsin, or papain at concentrations of 0.001, 0.005, or 0.01 g/100 g milk for 30 or 60 min to produce angiotensin-converting enzyme inhibitory and antioxidant peptides. Results showed that the proteolysis, antioxidant, and ACE-I activity gradually increased with the increase in the enzyme concentration and hydrolysis time. The protease-treated milk had the highest proteolytic and ACE-I activity, while the papain-treated milk had the lowest. The papain-treated milk exhibited the greatest Fe²⁺ chelating activity. The use of trypsin at concentration of 0.001 g/100 g milk for 60 min produced ACE-I and antioxidant activity without changes in the technological properties of milk.     

Casein peptides with inhibitory activity on lipid oxidation in beef homogenates and mechanically deboned poultry meat

Bioactive peptides obtained by enzymatic hydrolysis of casein may have antioxidant activity. In this work, casein peptides were obtained using the proteolytic enzymes Alcalase and Flavourzyme. Casein was hydrolyzed for 4 h at 50 °C and pH 8, and the resulting peptides were analyzed. The enzymatic hydrolysis with Flavourzyme resulted in higher concentration of soluble protein and free amino acids, and produced peptides with lower molecular mass than those obtained with Alcalase, as observed by gel permeation chromatography and polyacrylamide gel electrophoresis. Casein peptides obtained with Flavourzyme also exhibited greater antioxidant capacity using the ABTS radical method. Casein peptides (20 mg ml⁻¹) effectively inhibited lipid peroxidation in ground beef homogenates and mechanically deboned poultry meat. Casein peptides may be useful in meat processing as another naturally occurring antioxidant, helping to prevent off-flavor formation in meat products and increasing shelf life.     

Evaluation of the antioxidant activities and nutritional properties of ten edible plant extracts and their application to fresh ground beef

In this study, we assessed the antioxidant efficacy and nutritional value of 10 leafy edible plants and evaluated their potential as natural antioxidants for meat preservation. We measured total phenolic content, 2,2-diphenyl-1-picryl-hydrazil (DPPH) radical scavenging activity, and vitamin C, chlorophyll, and carotenoid contents of 70% ethanol and water extracts of the edible plants. Based on these results, we investigated the effects of butterbur and broccoli extracts on lipid oxidation in ground beef patties. Plant extracts and butylated hydroxytoluene (BHT) were individually added to patties at both 0.1% and 0.5% (w/w) concentrations. Thiobarbituric acid reactive substance (TBARS) values and color parameters were tested periodically during 12 days of refrigerated storage. TBARS levels were significantly lower (p ≤ 0.05) in the samples containing plant extracts or BHT than the non-treated control. In addition, the beef patties formulated with the selected plant extracts showed significantly (p ≤ 0.05) better color stability than those without antioxidants. These results indicate that edible plant extracts are promising sources of natural antioxidants and can potentially be used as functional preservatives in meat products.     

Effect of grape antioxidant dietary fiber on the lipid oxidation of raw and cooked chicken hamburgers

Efficiency of four concentrations (0.5, 1, 1.5 and 2%) of grape antioxidant dietary fiber (GADF) on susceptibility of raw and cooked chicken breast hamburger to lipid oxidation was investigated after 0, 3, 5 and 13 days of refrigerated storage at 4 °C. Color changes, sensorial qualities and acceptability by panellist were evaluated. Lipid oxidation was assessed by monitoring malondialdehyde formation with 2-thiobarbituric acid (TBARS) assay and radical scavenging capacity by ABTS method. A significant reduction in lightness and yellowness and a significant increase in redness as a result of GADF addition were observed in raw and chicken hamburgers. Addition of GADF significantly improved the oxidative stability and the radical scavenging activity in raw and cooked chicken hamburgers. The ability of GADF to prevent lipid oxidation was concentration-dependent. Acceptability of chicken meat was not affected by the addition of GADF. These results show that GADF is a very effective inhibitor of lipid oxidation and has potential as a natural antioxidant in raw and chicken cooked meats.     

The effect of ginger on diabetic nephropathy,plasma antioxidant capacity and lipid peroxidation in rats

Oxidative stress is a major factor in the pathogenesis of diabetic complications. We studied the effects of ginger powder on nephropathy induced by diabetes, and measured changes in plasma antioxidant capacity and lipid peroxidation.     

The triacylglycerol preparation of conjugated linoleic acid reduces lipid oxidation in irradiated, cooked ground beef patties

It is proposed that conjugated linoleic acid (CLA) would depress the lipid oxidation caused by irradiation of cooked, aerobically stored ground beef patties. The free fatty acid (FFA–CLA) and triacylglycerol (TAG–CLA) preparations of CLA were added at 0%, 1%, 2%, or 4% during the grinding process. Patties were irradiated at 1.5–2.0 kGy and frozen at −20 °C. Subsequently, the patties were tempered to 4 °C, cooked to 70 °C and held at 4 °C for 7 d. Enrichment of ground beef with CLA increased the cis-9,trans-11 and CLA trans-10,cis-12 CLA isomers in ground beef patties, even after cooking. Weight loss (P = 0.03) and percentage fat (P = 0.05) were higher in irradiated beef patties than in control patties. Irradiation decreased the concentration of α-linolenic acid (18:3n − 3) in the ground beef by over 60% (P = 0.07), whereas thiobarbituric acid reactive substances (TBARS) values were higher (P = 0.004) in irradiated beef patties than in control patties. The 1% concentration of added TAG–CLA reduced TBARS in irradiated ground beef patties, whereas 2% and 4% FFA–CLA depressed TBARS (CLA type × percentage interaction P = 0.04). Irradiation increased the cardboard and painty aromatic attributes (P  0.05), and FFA–CLA preparation increased the painty aromatic attribute and afterburn aftertaste, but these effects were not observed with the TAG–CLA preparation (CLA type × treatment interaction P < 0.04). Adding 1% TAG–CLA to ground beef during grinding can reduce lipid oxidation in irradiated, cooked ground beef patties without the negative aftertastes associated with the FFA–CLA preparation.     

Attenuated effects of peptides derived from porcine plasma albumin on in vitro lipid peroxidation in the liver homogenate of mice

Porcine plasma albumin was hydrolyzed with alcalase available for industrial application, and attenuated effects of peptides were evaluated using 4-Nitroquiunoline 1-oxide (4-NQO) as an inducing reagent. 4-NQO is a potent oral carcinogen, which has been found to induce lipid peroxidation in vivo and in vitro. Our results indicated that addition of 4-NQO resulted in increase of hydroxyl and superoxide radicals, glutathione peroxidase (GPx) activity and levels of malondialdehyde (MDA), and decrease in activities of superoxide dismutase (SOD) and catalase (CAT) and levels of glutathione (GSH). Simultaneous addition of peptides significantly attenuated lipid peroxidation. The lowermost molecular weight (MW) peptide fractions (<3 kDa) had the highest activity. This study also demonstrated that the attenuated effects of peptides might be due to the protective interactions between cells and peptides rather than the direct inhibition of 4-NQO by peptides. The results of this study showed the potential of utilizing porcine plasma albumin as a source of functional peptides.     

Effect of phosvitin on lipid and protein oxidation in ground beef treated with high hydrostatic pressure

In this study, we aimed to examine the effect of phosvitin on lipid and protein oxidation of raw and cooked ground beef treated with high hydrostatic pressure (HHP). Ground beef patty with 0, 500, or 1000 mg phosvitin/kg meat was treated with HHP at 0.1, 300, or 600 MPa. Half of the patties were used in a raw meat analysis, and the other half were used in a cooked meat analysis. Phosvitin and HHP treatment at 300 MPa synergistically reduced microbial growth, and HHP treatment at 600 MPa reduced microbial counts to undetectable levels (< 1 log CFU/g) throughout the length of the study in all samples. Phosvitin delayed lipid and protein oxidation in HHP-treated cooked and raw ground beef, respectively. However, phosvitin had no effect on the color changes of raw ground beef attributable to HHP. The results indicated that phosvitin could enhance the stability of lipids and proteins but not color changes of raw ground beef caused by HHP.     

Isolation and purification of a novel antioxidant protein from the water extract of Sundakai (Solanum torvum) seeds

Reactive oxygen species (ROS) at physiological concentrations may be required for normal cell function. Excessive production of ROS can be detrimental to cells, because ROS can cause oxidative damage to lipids, proteins, and DNA. Herein, we describe the isolation and purification of a novel antioxidant protein the water extract of dried, powdered Sundakai (Solanum torvum [Solanaceae]) seeds. Sundakai belongs to the Solanaceae family, a small shrub, which is distributed widely in India, Malaya, China, Phillipines and tropical America. Fifty percent of ammonium sulphate-precipitated crude water extract was fractionated on a Sephadex G100 column, which yielded two peaks, PI and PII. Peaks PI and PII inhibited lipid peroxidation up to 40% and 89%, respectively in linolenic acid micelles. Rechromatographing of peak PII on Sephadex G100 yielded a single peak, indicating the homogeneity of the purified protein. SDS–PAGE analysis indicated the molecular weight of the purified protein to be ∼28 kDa. The purified protein, at 0.8 μM, inhibited deoxyribose degradation induced by generation of hydroxyl radicals by 90% and scavenged DPPH (1,1-diphenyl-2-picrylhydrazyl) radicals by 76%. The reducing power and chelating power of the purified protein, at 0.8 μM, were found to be 72% and 85%, respectively. The protein, at 0.8 μM, also offered significant protection to calf thymus DNA damage induced by H₂O₂ (1 mM). Therefore, the present study demonstrates, for the first time, a novel protein from the water extract of Sundakai seeds as an excellent antioxidant.     

Packaging-specific influence of chitosan on color stability and lipid oxidation in refrigerated ground beef

We examined the influence of chitosan on lipid oxidation and color stability of ground beef stored in different modified atmosphere packaging (MAP) systems. Ground beef patties with chitosan (1%) or without chitosan (control) were packaged either in high-oxygen MAP (HIOX; 80% O₂ + 20% CO₂), carbon monoxide MAP (CO; 0.4% CO + 19.6% CO₂ + 80% N₂), vacuum (VP), or aerobic packaging (PVC) and stored at 1 °C. Chitosan increased (P < 0.05) redness of patties stored in PVC and CO, whereas it had no effect (P > 0.05) in HIOX. Chitosan patties demonstrated lower (P < 0.05) lipid oxidation than controls in all packaging. Control patties in PVC and HIOX exhibited greater (P < 0.05) lipid oxidation than those in VP and CO, whereas chitosan patties in different packaging systems were not different (P > 0.05) from each other. Our findings suggested that antioxidant effects of chitosan on ground beef are packaging-specific.     

Treatment of milk by gamma irradiation—effect of anoxia on lipid peroxidation and the survival of Pseudomonas aeruginosa

One of the causes of organoleptic changes in milk, lipid peroxidation, has been monitored by a sensitive assay newly applied to foodstuffs. It has been shown that even at medium radiation doses (up to 3600 Gy) negligible lipid peroxidation takes place, providing the milk is thoroughly gassed with nitrogen. Using a test spoilage microorganism, Pseudomonas aeruginosa (Schroeter), it was shown that the D₁₀ was 35 Gy in air and 105 Gy under anaerobic conditions. Dose/survival curves indicate that 5 decades of killing needed 200 Gy in air and 600 Gy under nitrogen. However, even after receiving a dose of 1600 Gy, both aerobically and anaerobically treated milk failed the methylene blue test at 21 days post-irradiation. The data indicate that, although the deleterious lipid peroxidation effects of irradiation can be minimised by the removal of air, this in turn results in a degree of protection being afforded to the bacteria, and that doses in excess of 1600 Gy are needed to effectively sterilise milk.     

Effects of plant extracts on microbial growth, color change, and lipid oxidation in cooked beef

The effects of butylated hydroxyanisole/butylated hydroxytoluene (BHA/BHT), grape seed extract (ActiVin), pine bark extract (Pycnogenol), and oleoresin rosemary (Herbalox) on microbial growth, color change, and lipid oxidation were investigated in cooked ground beef. When compared to the control, 1.0% ActiVin and Pycnogenol) effectively reduced the numbers of Escherichia coli O157:H7 and Salmonella Typhimurium, and retarded the growth of Listeria monocytogenes and Aeromonas hydrophila. Pycnogenol resulted in reductions of 1.7, 2.0, 0.8, and 0.4 log CFU/g, respectively, in numbers of E. coli O157:H7, L. monocytogenes, S. Typhimurium, and A. hydrophila, respectively, after 9 days of refrigerated storage. The color of cooked beef treated with ActiVin was less light (L*), more red (a*), and less yellow (b*) than those treated with BHA/BHT, Pycnogenol, and Herbalox. ActiVin and Pycnogenol effectively retained the redness in cooked beef during storage. The control showed significantly higher thiobarbituric acid-reactive substances (TBARS) and hexanal content over storage. BHA/BHT, ActiVin, Pycnogenol, and Herbalox retarded the formation of TBARS by 75%, 92%, 94%, and 92%, respectively, after 9 days, and significantly lowered the hexanal content throughout the storage period. Results of this work show that ActiVin and Pycnogenol are promising additives for maintaining the quality and safety of cooked beef.     

The combined effect of antioxidants and modified atmosphere packaging on protein and lipid oxidation in beef patties during chill storage

Effect of rosemary extract and ascorbate/citrate (1:1) in combination with modified atmosphere packaging (100% N(2), 80% O(2)/20% N(2)) on protein and lipid oxidation in minced beef patties during storage in the dark for up to 6 days at 4°C was investigated. A high level of oxygen in the packaging atmosphere was found to increase both lipid and protein oxidation during storage as evaluated by TBARS analysis of secondary lipid oxidation products and by 2,4-dinitrophenylhydrazine derivatization of protein carbonyls. Both antioxidant systems tested were found to inhibit lipid oxidation but not protein oxidation. In contrast, ascorbate/citrate was found to promote protein oxidation. Rosemary extract was found to regenerate or protect α-tocopherol whereas the packaging atmospheres had no effect on α-tocopherol stability. In high oxygen atmospheres both antioxidants protected the fresh red meat colour with ascorbate/citrate being more efficient than the rosemary extract, whereas no effect of antioxidant on meat colour was found in beef patties stored in 100% nitrogen.     

Identification of antioxidant peptides derived from egg-white protein and its protective effects on H2O2-induced cell damage

To identify novel antioxidant peptides from egg-white protein and investigate antioxidant mechanism, the hydrolysate of egg-white protein was purified by ultrafiltration and a Sephadex G-15 column. The peptides VYLPR, EVYLPR, VEVYLPR and VVEVYLPR were identified from the fraction with the highest antioxidant activity. The Results showed that the peptide VYLPR exhibited the strongest protective effect on HEK-293 cells. The viability of cells recovered to 97.45 ± 1.98% with pre-treatment of 20 μm VYLPR. We further investigated antioxidant mechanism. The results showed VYLPR could inhibit lipid peroxidation process, maintain cell membrane integrity, inhibit intracellular LDH activity, reduce MDA content, and improve the activity of antioxidant enzyme T-SOD and GSH-Px. This work could help researchers in understanding antioxidant mechanism of peptides and also contribute to developing functional egg-white product.     

Structural characteristics of peptides extracted from Cantonese sausage during drying and their antioxidant activities

Two peptide fractions (P1 and P2) were isolated from Cantonese sausages at different drying periods by ultrafiltration. P1 possessed a stronger 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in the first 18 h of drying and a weaker activity during the rest period, comparing with P2. The analysis of amino acid composition indicated that histidine was the major amino acid in both peptide fractions, which content was changed during the process. In addition, Glu, Gly, Arg were important constituent amino acids in both peptide fractions. A significant (P < 0.05) decrease of >10 kDa peptide fraction and increases of 2–3 and <2 kDa peptide fractions were detected when comparing with peptides at 0 h. However, no significant difference was found for all the peptide fractions at different drying periods, except for 2–3 kDa peptides. A significant (P < 0.05) correlationship was found between lipid oxidation and antioxidant activities.

Industrial relevance

Cantonese sausage has gained much popularity and acceptance in China and many other countries. In this work, two peptide fractions were isolated from Cantonese sausage. A good antioxidant activity was observed for each peptide fraction. Endogenous peptides resulting from proteolysis offer a promising approach to improve lipid stability for meat industry. It is helpful to retard lipid oxidation in order to improve its quality.     

Evaluation of cholesterol and lipid oxidation in raw and cooked minced beef stored under oxygen-enriched atmosphere

Oxygen-enriched modified atmosphere packaging (MAP) represents an important means to stabilize meat colour but may lead to an increase in lipid oxidation, influencing the acceptability and safety of the product. In this work, the effect on cholesterol and lipid susceptibility to oxidation was investigated in commercial minced beef held under MAP (80% O(2)/20% CO(2)). Cholesterol oxidation products (COPs), peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) were determined, before and after pan frying, at 1, 8 and 15 days since packaging under refrigerated storage (3-4°C). 7α-Hydroxycholesterol, 7β-hydroxycholesterol and 7-ketocholesterol were the more abundant COPs identified. COPs significantly increased in raw beef during storage: after 1, 8 and 15 days since packaging COPs were at the levels of 10.4, 30.7 and 60.5μg/g of fat, respectively. Cooking did not affect cholesterol oxidation in freshly packaged minced beef but led to a rise in COPs amount with respect to raw muscle after 8 and 15 days of storage. The trend in cholesterol oxidation reflected the progressive increase in lipid peroxidation rate brought by MAP conditions.     

A review of natural antioxidants and their effects on oxidative status, odor and quality of fresh beef produced in Argentina

Meat derived from pasture feeding, is associated with a high level of antioxidants. Antioxidants are incorporated within cell membranes and protect tissues against oxidation from reactive oxygen species. This maintains the overall quality of meat and secondary products. This paper reviews the implications of incorporating natural antioxidants into fresh beef, focusing on the benefits of feeding cattle good quality pasture. Pasture samples typically have higher levels of α-tocopherol, β-carotene, ascorbic acid and glutathione than feedlot samples. These compounds retard lipid and protein oxidation in fresh and stored meat, and preserve the color and odor quality of beef. The significance of antioxidant enzymes is variable, because their behavior depends on individual redox status before slaughter. Understanding total antioxidant activity requires information on antioxidant and pro-oxidant status. With an abundance of pasture, Argentina has a natural advantage in producing meat with a high antioxidant value.     

Secondary structures and their effects on antioxidant capacity of antioxidant peptides in yogurt

The pH, thermal, urea, and trifluoroethanol treatments were used to denature yogurt antioxidant peptides, and their antioxidant capacities and secondary structures were analysed. Results showed that yogurt peptides treated at pH 6.0 or above 50°C exhibited higher antioxidant activity than the control, whereas trifluoroethanol treatment resulted in a reduction and urea treatment held invariability. Peptide secondary configuration analyses revealed that these treated peptides significantly decreased their α-helical conformation but enhanced the β-sheet presence as well as an obvious alteration on β-turn and random coil structures. These peptide conformation alterations contributed to their antioxidant property increase.