Fractionation and characterization of tartary buckwheat flour proteins

Protein fractions (albumin, globulin, prolamin and glutelin) were extracted from defatted tartary buckwheat flour. Albumin was the predominant protein fraction (43.8%) followed by glutelin (14.6%), prolamin (10.5%), and globulin (7.82%). Albumin was relatively rich in histidine, threonine, valine, phenylalanine, isoleucine, leucine and lysine. Globulin had high levels of methionine and lysine. Prolamin was high in histidine, threonine, valine, isoleucine, and leucine. Glutelin was rich in histidine, threonine, valine, isoleucine, and leucine. SDS–PAGE analysis, under non-reductive and reductive conditions, showed that disulfide bonds existed in the four fractions. Non-reduced albumin showed major bands at 64, 57, 41, and 38 kDa, and globulin at 57, 28, 23, 19 and 15 kDa. Reduced albumin and globulin shared two common bands at 41 and 38 kDa. Reduced prolamin showed major bands at 29, 26, 17 and 15 kDa. In vitro pepsin digestibility of the four fractions (from high to low) was: albumin > globulin > prolamin and glutelin.     

Preservation of the endogenous antioxidant system of fish muscle by grape polyphenols during frozen storage

The capacity of a phenolic extract, OW, obtained from grape (Vitis vinifera) by-products, and of a purified fraction of procyanidins from OW, fraction IV, for preservation of endogenous antioxidants of fatty fish was investigated during frozen storage. They were used in muscle concentrations of 0.01% (w/w). Grape polyphenols were compared with propyl gallate, a synthetic antioxidant. The exogenous compounds were added to minced mackerel (Scomber scombrus) muscle and horse mackerel (Trauchurus trauchurus) fillets, before freezing at –10 °C. The results demonstrated that grape polyphenols and propyl gallate inhibit the depletion of endogenous -tocopherol, ubiquinone-10 and total glutathione. Grape polyphenols and propyl gallate showed similar efficiency for preservation of ubiquinone, in both minced and filleted muscle, and total glutathione, in minced muscle. Total glutathione in the fillets was better maintained by propyl gallate than grape polyphenols. The endogenous antioxidant more efficiently preserved by grape polyphenols and propyl gallate was -tocopherol. Its loss elapsed faster in the order control> OW>fraction IV>propyl gallate. The depletion of -tocopherol was highly correlated with the evolution of lipid oxidation. The development of lipid oxidation was repressed, while the concentration of -tocopherol was not reduced up to critical levels.     

Effect of actinidin from kiwifruit (Actinidia deliciosa cv. Hayward) on the digestion of food proteins determined in the growing rat

This study aimed to determine the effect of dietary actinidin (provided as Hayward kiwifruit) on the gastric and small intestine digestion of six food protein sources in rats. For each protein source, two semi-synthetic test diets were formulated containing either freeze-dried Hayward kiwifruit (actinidin present) or freeze-dried Hort16A kiwifruit (actinidin absent). Actinidin activity is extremely low in Hort16A kiwifruit. Titanium dioxide was also included as an indigestible marker. Rats were fed freshly-prepared diets, euthanised and the gastric and ileal contents collected. The chyme and digesta samples were subjected to electrophoresis (SDS–PAGE), densitometry and titanium analysis and the degradability of individual proteins calculated. Dietary actinidin had no (p > 0.05) effect on the gastric degradability of zein and whey protein isolate but increased gastric degradability of beef muscle protein, gelatin, soy protein isolate and gluten by 40%, 60%, 27% and 29% units, respectively. Dietary actinidin had little or no effect on ileal protein degradability. Overall, dietary actinidin enhanced the gastric digestion of some food proteins.     

Catalytic mechanisms of metmyoglobin on the oxidation of lipids in phospholipid liposome model system

The catalytic mechanism of metmyoglobin (metMb) on the development of lipid oxidation in a phospholipid liposome model system was studied. Liposome model system was prepared with metMb solutions (2.0, 1.0, 0.5, and 0.25 mg metMb/mL) containing none, diethylenetriamine pentaacetic acid (DTPA), desferrioxamine (DFO), or ferric chloride and lipid oxidation was determined at 0, 15, 30, 60, and 90 min of incubation at 37 °C. Metmyoglobin catalysed lipid oxidation in the liposome system, but the rate of lipid oxidation decreased as the concentration of metMb increased. The amount of free ionic iron in the liposome solution increased as the concentration of metMb increased, but the rate of metMb degradation was increased as the concentration of metMb decreased. The released free ionic iron was not involved in the lipid oxidation of model system because ferric iron has no catalytic effect without reducing agents. Both DFO and DTPA showed antioxidant effects, but DFO was more efficient than DTPA because of its multifunctional antioxidant ability as an iron and haematin chelator and an electron donor. The antioxidant activity of DTPA in liposome solution containing 0.25 mg metMb/mL was two times greater than that with 2 mg metMb/mL due to the increased prooxidant activity of DTPA-chelatable compounds. It was concluded that ferrylmyoglobin and DTPA-chelatable haematin generated from the interaction of metMb and LOOH, rather than free ionic iron, were the major catalysts in metMb-induced lipid oxidation in phospholipid liposome model system.     

Lipid oxidation in salted-dried fish: II. The effect of photosensitisers on the rate of oxidation of a fish oil

The contribution of photosensitised oxidation to the extent of lipid oxidation of a model fish oil system has been considered using a number of methods of measurement, namely peroxide value, polyene index and the initial rate of oxygen uptake. This represents the initial stages of the development of a model system which will eventually simulate the processing and storage of salted, sun-dried fish. The effects of the photosensitisers, protoporphyrin IX, riboflavin and myoglobin on the rate of oxidation of a highly unsaturated fish oil in a model system, have been studied under dark or photosynthetic light conditions at 30°C. As has previously been established, the initial rate of oxygen uptake proved to be a more reliable method than the peroxide value or the polyene index in determining the extent of lipid oxidation of the oil in the model system. Protoporphyrin IX, riboflavin and myoglobin all increased the rate of oxygen uptake significantly in photosynthetic light at 30°C, from a rate of 0–158 μl O₂g^?1for the control to 0.335 μl O₂g^?1min^?1, 0.308 μl O₂ g^?1min^?1 and 0.461 μl O₂g^?1min^?1, respectively. However, in the dark at 30°C their addition had no effect on the initial rate of oxygen uptake, showing that these components increased the rate via a photosensitised oxidation mechanism rather than by autoxidation. The addition of β-carotene, a well-established singlet oxygen quencher, to the systems containing the photosensitisers at 30°C in the light significantly reduced the rate of oxygen uptake, but not to the same rate as in the control. Photosensitised oxidation has been classified as Type I or Type II; the reduction observed in oxygen uptake rate in the presence of β-carotene shows the Type II mechanism involving singled oxygen to be important. Studying β-carotene on its own in the model system at 30°C showed an unexpected increase in the rate of oxygen uptake both in the light and in the dark.     

Effects of xylooligosaccharides and sugars on the functionality of porcine myofibrillar proteins during heating and frozen storage

The gel properties of mixed gels of salt-soluble proteins (SSP) and xylooligosaccharides (XOS) or other sugars were investigated. Furthermore, the water-holding capacity, synaeresis, and protein surface hydrophobicity of heated SSP/sugar mixed gels under frozen storage were evaluated. Addition of XOS and sugars lowered expressible moisture and thus elevated the water-holding capacity of mixed gels. The protein surface hydrophobicity (RSo) of heated (71.1 °C) XOS-added gels was lower than that of other treatments, indicating possible lower exposure of protein hydrophobic groups. Results from freeze/thaw stability and protein solubility showed a descending pattern of SSP/sugar mixed gels during frozen storage, with XOS-added gels being better in both test items than other treatments. Increasing saccharide levels improved freeze/thaw stability and protein solubility of frozen mixed gels. Addition of xylooligosaccharides resulted in the lowest RSo among treatments and appears to impact better cryoprotective function to meat proteins than other sugars.     

Influence of Botrytis cinerea infection on Champagne wine proteins (characterized by two-dimensional electrophoresis/immunodetection) and wine foaming properties

Proteins are implicated in the foam stabilization of Champagne wines. They may have a grape, yeast, bacteria or fungal origin. Botrytis cinerea is a widespread fungal pathogen, which is the causal agent for gray mold. The first part of this work showed the deleterious effect of the presence of this fungus on the foaming properties of a champenois base wine. Foamability and foam stability were reduced, respectively, by 47.7% and 33.3% in the botrytized wine, as compared to the healthy wine. In a second part, SDS–PAGE and two-dimensional electrophoresis (2-DE), coupled with immunodetection, were used to study (thoroughly) the protein patterns of both wines. With 2-DE and silver-staining detection, the disappearance of numerous spots, located in an acidic pH range, was observed. Indeed, the number of spots detected was about two times more abundant in the healthy wine than in the botrytized one, suggesting that a proteolysis occurred. On the other hand, the presence of new proteins, likely fungal proteins, proteins secreted by the plant as a response to B. cinerea infection, or even protein fragments resulting from partial proteolysis, was detected in the botrytized wine. All these modifications of the wine protein content were undoubtedly due to the presence of B. cinerea and this might be a reason for the loss of foaming properties of Champagne base wines, though no relationship between these two phenomena can be established from the results obtained.     

Functional and nutritional characteristics of proteins and lipids recovered by isoelectric processing of fish by-products and low-value fish: A review

Several fisheries are over-exploited and may collapse; yet the amounts of fish processing by-products containing muscle proteins and ω-3-rich oil are staggering. The by-products are land-filled, ground and discarded or otherwise diverted from human consumption. Due to the lack of technology to recover proteins and lipids from by-products or low-value species, this tremendous resource is unavailable for human consumption. Isoelectric solubilisation/precipitation (ISP) allows efficient recovery of fish proteins and oil which retain functionality and nutritional value of food products. Isoelectric point (pI) is a pH where protein maintains zero electrostatic charge. At pI, protein–protein hydrophobic attraction overcomes protein–water electrostatic attraction resulting in isoelectric precipitation. Conversely, isoelectric solubilisation occurs at a pH different from pI, whereby protein–water attraction and protein–protein electrostatic repulsion are favoured. Therefore, protein solubility/insolubility is induced by ISP, respectively. Consequently, ISP allows selective protein recovery. Lipids are also recovered during ISP processing. This article reviews recent ISP developments to recover proteins and lipids from by-products and low-value marine species.     

Effect of bleeding treatment and perfusion of yellowtail on lipid oxidation in post-mortem muscle

The present study investigated the effects of bleeding treatment and perfusion of antioxidant compounds on lipid oxidation in ordinary and dark muscles of yellowtail in the early stage of ice storage. The lipid hydroperoxide contents of dark muscles obtained from yellowtails with and without bleeding treatment were higher and increased more rapidly than those of ordinary muscles. There were no significant differences in the rates of change of the lipid hydroperoxide content (up to 48 h), fatty acid composition and metmyoglobin formation between dark muscles with and without bleeding treatment. Physiological saline containing ascorbic acid or 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox^®) was perfused into live yellowtail or added to minced dark muscle. Trolox^® significantly (P < 0.01) delayed the accumulation of lipid hydroperoxide in dark muscle compared to ascorbic acid in perfusion experiment. These results indicate that simply removing a portion of the blood from live yellowtail by bleeding is not sufficient to prevent lipid oxidation in the early stage of ice storage. Contrary to this, addition of antioxidants into fish flesh is effective to delay lipid oxidation in post-mortem muscle.     

Effect of lipid oxidation and frozen storage on muscle proteins of Atlantic mackerel (Scomber scombrus)

The effect of storage on the lipids and proteins in Atlantic mackerel stored for up to 24 months at ?20 and ?30 °C was studied. Traditional methods including the peroxide value, thiobarbituric acid‐reactive substances (TBARS) and a reverse phase HPLC method were used to determine the primary and secondary lipid oxidation products. All tests showed an increase in lipid oxidation products with storage time and at a higher storage temperature of ?20 °C compared with samples stored at ?30 °C. Antioxidants had a significant effect (P < 0.01) on the inhibition of lipid oxidation, as shown by the reduction in peroxide value and hydroxides, and malondialdehyde formation. Similarly, deterioration of protein structure and functionality in mackerel stored for 3, 6, 12 and 24 months was greater at ?20 than ?30 °C. ATPase activity in the myosin extract of Atlantic mackerel showed a significant decrease (P < 0.01) with progressive frozen storage. Protein solubility in high salt concentration (0.6 M NaCl) decreased (P < 0.01) during storage at both ?20 and ?30 °C but was greater at ?20 °C. Interestingly, antioxidants BHT, vitamin C and vitamin E protected the proteins against complete loss of ATPase activity and protein solubility to a significant level (P < 0.01) for up to 1 year at ?20 °C compared with samples stored without antioxidants. This study confirms the deleterious effect of lipid oxidation products on protein structure and function in frozen fatty fish. © 2002 Society of Chemical Industry     

Lipid oxidation in salted-dried fish: The effect of temperature and light on the rate of oxidation of a fish oil

In relation to studies of lipid oxidation during the processing and storage of salted sun-dried fish, the measurement of the initial rate of oxygen uptake has been studied on a model system consisting of a highly polyunsaturated fish oil. This parameter has been used for assessing the effects of temperature and light conditions on the model system and has been compared to changes in the peroxide value and the polyene index. Assessment of oxygen uptake using a Gilson respirometer was found to give a clearer indication than other methods of assessment of the effect of different conditions on fish oil oxidation. At 30°C the initial rate of oxygen uptake increased from 0-103 μl O₂, g^?1 miir^?1 to 0-158 μl O₂, g^?1 min^?1 with a change in light conditions from dark to photosynthesis light. At 40°C, the rate of oxygen uptake was fester than at 30 °C, and the effect of light was more pronounced, giving values of 0.188 μl O₂, g^?1 min^?1 and 0.483 μ O₂, g^?1 min^?1, for dark and photosynthesis light, respectively. The observed results for the measurement of the initial rate of oxygen uptake suggest its use as a sensitive and reliable method for the investigation of the contribution of the many components in salted sun-dried fish towards the rate of oxidation and subsequent rancidity.     

鱼肉质构的影响因素及测定方法研究进展

鱼类产品是居民膳食重要的动物性蛋白源, 而质构特征是评价鱼肉品质的关键指标。为系统总结分析影响鱼肉质构的主要因素, 本文首先分析了鱼肉质构与肌纤维、结缔组织及肌内脂肪的内在关系; 在此基础上, 从养殖方式、饲料配方、捕获季节及处死方式等角度综述了影响鱼肉质构的外在因素, 并分析了内源蛋白酶降解、细胞冷冻冰晶损伤及蛋白冷冻变性在低温贮藏鱼肉质构软化中的作用; 最后总结了鱼肉质构的仪器测定方法, 旨在为鱼类产品食用品质评价及控制提供理论参考。     

Effect of high pressure on the calpain-calpastatin system in fish muscle

ABSTRACT:  Calpains (calcium-activated neutral proteases) of sea bass ( Dicentrarchus labrax L.) muscle may participate in the degradation of muscle tissue during postmortem storage. These enzymes are regulated by calpastatin, their endogenous specific inhibitor. The objective of this study was to evaluate the changes encountered by the calpain system during the postmortem storage of fish muscle after high-pressure treatment. From 100 MPa, high-pressure treatment of purified calpains results in a loss of their activity as well as in the dissociation of the heterodimeric form. In muscle, the high-pressure processing decreases the initial activity of calpain. This loss in activity may be due to an inactivation by a change of structure. Initial calpastatin activity is not modified by the high-pressure treatment, but it decreases during the storage from the beginning for a treatment at 300 MPa after which calpastatin is stable during 2 d. Therefore, this study also suggests that high-pressure treatment could be a useful way to improve fish flesh quality.     

Anti-oxidant activity of added tea catechins on lipid oxidation of raw minced red meat, poultry and fish muscle

The comparative anti-oxidative effects of added tea catechins (TC) and α-tocopherol to raw minced red meat (beef and pork), poultry (chicken, duck and ostrich) and fish (whiting and mackerel) muscle on susceptibility to lipid oxidation were investigated during 10 days of refrigerated (4 °C) display. Fresh meats, poultry and fish, purchased from a local market, were trimmed to remove bones, skin and surface fat and minced through a 4 mm plate. The minced muscle of each species was treated with either the addition of 300 mg TC kg^–1 minced muscle (TC300) or 300 mg α-tocopherol kg^–1 minced muscle (VE300). Minced muscle without any additives served as control (C). Oxidative stability (TBARS) was measured at 3-day intervals. Total lipids, fatty acid composition, total iron and haem iron from minced muscle for each species were also analysed. The susceptibility of untreated minced muscle to lipid oxidation was in the decreasing order: mackerel > beef > duck > ostrich > pork ≥ chicken > whiting. This may be because of the different content of total fat, iron and fatty acid composition between species. The TC300 significantly ( P  < 0.05) reduced lipid oxidation compared with controls for all seven species as shown by lower TBARS values. The anti-oxidant potential of TC was two to fourfold greater than that of α-tocopherol at the same concentration and this potential was species dependent. The VE300 showed limited capacity in inhibiting lipid oxidation for pork, chicken, duck and whiting. The results obtained show that TCs are powerful natural antioxidants when used in minced muscle food.     

Effect of different cooking methods on lipid oxidation and formation of free cholesterol oxidation products (COPs) in Latissimus dorsi muscle of Iberian pigs

The aim of this work was to study the influence of different cooking methods (grilled (GR), fried (FP), microwave (MW) and roasted (RO)) on lipid oxidation and formation of free cholesterol oxidation products (COPs) of meat from Iberian pigs that have been fed on an intensive system. Moisture and total lipid content, TBARs, hexanal and COPs were measured in Latissimus dorsi muscle samples. Cooking did not produce changes in total lipid content in meat but induced significantly higher lipid oxidation (TBARs and hexanal values) (p < 0.001) and cholesterol oxidation (COPs) (p < 0.01). When the different cooking methods were studied, the grilled method was the least affected by lipid oxidation (TBARs and hexanal) compared to the others. There were no significant differences among different cooking methods on COPs values. The most abundant cholesterol oxides were both 7α-hydroxycholesterol and 7β-hydroxycholesterol in all groups studied.     

Emulsifying and foaming properties of Phaseolus vulgaris and coccineus proteins

Protein isolates from two Phaseolus cultivars, common bean (Phaseolus vulgaris L.) and scarlet runner bean (Phaseolus coccineus L.), were prepared by wet extraction methods (isoelectric precipitation – 4000 rpm, ultrafiltration, extraction with NaCl 2%, and isoelectric precipitation – 9900 rpm). The protein isolates were characterized by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and then evaluated for their solubility. The emulsion stability of emulsions produced at pH 7.0 and 5.5 with 1% or 2% or 3% w/v protein isolate was evaluated by average droplet size diameter, viscosity and creaming measurements. Emulsions with 1% protein content were unstable through storage. Emulsions with 3% w/v protein isolate concentration, extracted by ultrafiltration at pH 5.5 from both cultivars, were flocculated; this was more pronounced for coccineus isolates. The foaming properties, for the respective foams, were investigated. Foams with 1% w/v protein showed little foaming ability Ultrafiltration isolates produced more foam, which was especially stable at pH 5.5.     

Effect of heat,rutin and disulfide bond reduction on in vitro pepsin digestibility of Chinese tartary buckwheat protein fractions

Protein fractions (albumin, globulin, prolamin and glutelin) were extracted from defatted tartary buckwheat flour. The in vitro pepsin digestibilities of the four protein fractions were different, and albumin was more susceptible to pepsin hydrolysis. The native structure of the four protein fractions may be destroyed by heat treatment, and the digestibilities were all improved significantly (P < 0.05). Adding rutin to the digestion mixture of the four fractions did not cause a decrease in pepsin digestibility, although it did cause a significant increase in certain instances (P < 0.05). Treatment with β-mercaptoethanol (2-ME) only caused a higher initial proteolysis rate and did not increase the final digestibility distinctly except for prolamin. After pepsin digestion, the remaining proteins of unhydrolyzed albumin, globulin, prolamin and glutelin (untreated) shared some similarities. They also exhibited a minor band at 20,000 Da and a broad band at 10,000–14,000 Da.     

深度油炸过程煎炸油的氧化及其对油炸外裹糊鱼块品质的影响

目的 探究深度油炸过程煎炸油的氧化,并进一步研究煎炸油氧化对油炸外裹糊鱼块品质的影响。方法 分别采用棕榈油、大豆油、葵花籽油、小麦胚油在150、160、170、180、190℃下油炸外裹糊鱼块,测定煎炸油的酸价、过氧化值、黏度、介电常数以及油炸外裹糊鱼块外壳的水分含量、油脂含量、表面色度。结果 随着油炸温度的升高,煎炸油的游离脂肪酸含量增加,导致煎炸油的酸价、黏度和介电常数升高,过氧化值呈现波动下降的趋势;油炸外裹糊鱼块的水分含量逐渐减少,油脂含量逐渐增加,L*和b*呈递减趋势、a*呈递增趋势,且使用4种煎炸油的各项指标存在明显差异。结论 煎炸油的油炸温度和不饱和脂肪酸含量显著影响了煎炸油的氧化,导致外裹糊鱼块深度油炸过程中水分蒸发和油脂吸收有明显差异,最终影响了油炸外裹糊鱼块的品质。     

Effect of frozen storage on lipids and lipolytic activities in the longissimus dorsi muscle of the pig

 Samples of longissimus dorsi muscle from pigs were vacuum-packed and stored at –18  °C for a 6-month period. The quantity of total lipids, non-polar lipids, phospholipids and cholesterol remained unchanged during storage. However, there was a decrease (1.4%) in the polyunsaturated fatty acid percentage of the phospholipid fraction after 6 months of frozen storage, mainly due to the decrease in linoleic fatty acid. Nevertheless, there was no change in fatty acid composition of the non-polar lipid fraction. Phosphatidylethanolamine was the phospholipid most affected during the frozen storage, with a significant decrease from an intial percentage of 26.6% to 23.0% after 6 months of storage. Important activities of muscle lipolytic enzymes were still recovered after the storage, which explains the continuous release of free fatty acids reported during the process, with a net increase of 50.6 mg/100 g dry matter. The highest release of free fatty acids was reported during the 1st month of frozen storage. At the 6th month of frozen storage the compositions of both the free fatty acid and phospholipid fractions were similar. With respect to oxidation, the thiobarbituric acid test number showed a slight increase during the process while the peroxide value remained unchanged. Received: 16 March 1998 / Revised version: 17 June 1998