Structural modification of soy protein by 13-hydroperoxyoctadecadienoic acid

To reveal the role of primary products of lipid peroxidation during soy protein oxidation process, oxidative modification of soy protein by 13-hydroperoxyoctadecadienoic acid (13-HPODE) generated by lipoxygenase-catalyzed oxidation of linoleic acid was investigated in this article. Incubation of soy protein with increasing concentration of 13-HPODE resulted in generation of protein carbonyl derivatives and loss of protein sulfhydryl groups. Circular dichroism spectra indicated that exposure of soy protein to 13-HPODE led to loss of α-helix structure. Effect of oxidation on tertiary structure was demonstrated by surface hydrophobicity and tryptophan fluorescence. Surface hydrophobicity gradually decreased, accompanied by loss and burial of some tryptophan residues. The results of surface hydrophobicity and tryptophan fluorescence implied that aggregation was induced by oxidation. Size exclusion chromatogram indicated that the extent of aggregation was increased in a 13-HPODE dose-dependent manner. Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated that non-disulfide linkages were involved in aggregate formation, and β-conglycinin was more vulnerable to 13-HPODE than glycinin.     

Structural modification of soy protein by the lipid peroxidation product malondialdehyde

BACKGROUND: Protein oxidation results in covalent modification of structure and deterioration of functional properties of target protein. Oxidation extent of soy protein was affected by the content and type of lipid peroxidation (LPO) products in defatted soybean flours during storage and processing. Malondialdehyde (MDA) was selected as a secondary byproduct of LPO to investigate the effects of oxidative modification of LPO‐derived reactive aldehyde on soy protein structure. RESULTS: MDA reacted with ε‐amino and sulfhydryl groups of soy protein, and resulted in an increase in protein carbonyl groups but a decrease in sulfhydryl/disulfide, free amines and lysine. The decrease in solubility, surface hydrophobicity and intrinsic fluorescence indirectly indicated that MDA induced soy protein aggregation, and results of high‐performance size‐exclusion chromatography directly showed that gradual aggregation of soy protein was induced by increasing concentration of MDA. Results of electrophoresis indicated that MDA caused soy protein aggregation, and non‐disulfide covalent bonds were involved in aggregate formation. CONCLUSION: The results showed that sensitivity of soy protein was related to MDA concentration. Soy protein gradually aggregated with increase of MDA concentration; β‐conglycinin was more sensitive to MDA modification than glycinin. Copyright © 2009 Society of Chemical Industry     

Oxidative modification of soy protein by peroxyl radicals

Oxidative modification of soy protein by peroxyl radicals generated in a solution containing 2,2’-azobis (2-amidinopropane) dihydrochloride (AAPH) under aerobic condition was investigated. Incubation of soy protein with increasing concentration of AAPH resulted in gradual generation of protein carbonyl derivatives and loss of protein sulphydryl groups. Circular dichroism spectra indicated that exposure of soy protein to AAPH led to loss of α-helix structure. Effect of oxidation on tertiary structure was demonstrated by surface hydrophobicity and tryptophan fluorescence. Surface hydrophobicity steadily decreased, accompanied by loss and burial of some tryptophan residues, indicating that soy protein gradually aggregated. The results of the size exclusion chromatogram (SEC) implied that incubation caused an AAPH-dose-dependent increase of fragmentation and aggregation of oxidised soy protein. Sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) indicated that non-disulphide linkages were involved in aggregate formation, and β-conglycinin was more vulnerable to peroxyl radicals than glycinin.     

Effects of oxidative modification on thermal aggregation and gel properties of soy protein by peroxyl radicals

Effects of protein oxidation on thermal aggregation and gel properties of soy protein by 2,2′‐azobis (2‐amidinopropane) dihydrochloride (AAPH)‐derived peroxyl radicals were investigated in this article. Incubation of soy protein to increase concentration of AAPH resulted in a decrease in particle size and content of thermal aggregates during thermal‐induced denaturation. Protein oxidation resulted in a decrease in water‐holding capacity (WHC), gel hardness and gel strength of soy protein gel. An increase in coarseness and interstice of the gel network was accompanied by uneven distribution of interstice as extent of oxidation of soy protein increased. A decrease in disulphide content and formation of oxidation aggregates in the process of oxidative modification were contributed to the decline of particle size and content of thermal aggregates during thermal‐induced denaturation, leading to a decrease in WHC, gel hardness and gel strength of soy protein gel.     

Inhibition of protein and lipid oxidation by rapeseed, camelina and soy meal in cooked pork meat patties

In this study protein-containing by-products of deoiling processes rich in phenolics were applied to meat to be used as potential food ingredients in developing meat products with antioxidant effect. The effect of rapeseed meal (Brassica rapa L.), camelina meal (Camelina sativa), soy meal and soy flour (from soybean, Glycene max L.), in inhibiting oxidation of lipids and proteins was tested in cooked pork meat patties. A commercial CO₂ extract from rosemary (Rosmarinus officinalis) was used as a reference material alone and in combination with the other plant materials. The cooked pork meat with added plant materials was oxidized for 10 days at 5 °C under light. The oxidation was followed by measuring the formation of hexanal, pentanal and propanal by headspace gas chromatography and the formation of protein carbonyls by converting them to 2,4-dinitrophenylhydrazones (DNPH). Rapeseed meal (0.5 and 0.7 g/100 g meat) and camelina meal (0.7 g/100 g meat) as such and their combination (addition of 0.5 g/100 g) with rosemary extract (0.04 g/100 g) were effective antioxidants toward both protein and lipid oxidation while soy meal and flour were effective only in combination with rosemary extract.     

Structural and antioxidant modification of wheat peptides modified by the heat and lipid peroxidation product malondialdehyde

Wheat peptides, the biological active peptides derived from foods, has an array of biological actions, including antiobesity, antimicrobial, and angiotensin I-converting enzyme inhibitory effects in mammalian species. Recent studies showed that some wheat peptides may show the noteworthy antioxidant potency against the peroxidation of lipids or fatty acids, but the effect of oxidation on its antioxidant activities is unclear. In the present study, we demonstrate that heat and malandialdehyde (MDA)-oxidized wheat peptides lose its surface hydrophobicity and reducing power, and show a relatively lower free radical-scavenging activitiy in vitro. Those modifications also lead to gradual formation of aggregates in wheat peptides and induce more reactive oxygen species (ROS) production in vivo. These findings indicate that oxidation may influence the functional properties and directly alter the structure of wheat peptides, and lead to the loss of its antioxidant potency both in vitro and in vivo, thereby providing a novel explanation for some of the potential health risks proposed for oxidized food in human.     

Antioxidant effects of soy sauce on color stability and lipid oxidation of raw beef patties during cold storage

This study was conducted to evaluate the antioxidant effects of soy sauce on lipid oxidation and color stability of raw beef patties. Raw beef patties were formulated with four solutions such as NaCl (sodium chloride solution), NaCl/SS (1:1 ratio of sodium chloride and soy sauce solution), SS (soy sauce solution), or SS/A (soy sauce solution combined with 0.05% ascorbic acid) in the same salt concentration. Addition of soy sauce resulted in the decreased pH, lightness, and increased yellowness. Treatment SS/A had the lowest percent of metmyoglobin during storage (P < 0.05). A reduction (P < 0.05) in the 2-thiobarbituric acid, peroxide, and conjugated diene concentration as result of soy sauce addition were observed in treatments SS and SS/A at the end of the storage period. There were no differences (P > 0.05) in free fatty acid concentration at the end of storage. The combined addition of soy sauce and ascorbic acid greatly improved (P < 0.05) color stability and retarded lipid oxidation.     

Soft cheese-like product development enriched with soy protein concentrates

The objective of this work was to develop spreadable cheese-like product from skimmed cow's milk, supplemented with soybean proteins. The proteins were concentrated by ultrafiltration, and then freeze-dried. The cheese-like products were prepared using the acid coagulation method, without maturation, obtaining smooth products with creamy texture. The samples were identified as low-fat cheese, being the fat content between 10 and 13 (g/100 g), with moisture in the range of 68.07–70.75 (g/100 g). Compared to a control cheese, the samples containing soy protein concentrate, showed an increase of 6.8–17 in proteins and 22–32 (g/100 g) in fats; also the yield increased. The viscoelastic behavior of the samples was analyzed using oscillatory dynamic tests; in all the samples G′ was higher than G″. Texture, surface color and microstructure of the cheese samples were determined. Microbial analysis showed that the incorporation of potassium sorbate increased more than twice the shelf-life of the products (up to 60 days) in comparison to the samples without preservative. Furthermore, the products were accepted by a sensory panel. Considering that both, proteins and fats, are of vegetable origin with high biological value and unsaturated fats, the developed cheese-like products were considered as functional foods.     

脂质氧化产物-蛋白质相互作用研究进展

富含不饱和脂质的食品及其原料在加工和储存过程中易受多种因素影响而发生氧化。在此过程中生成的过氧化自由基和其他次级氧化产物能够使得蛋白质发生复杂变化。脂质氧化产物与蛋白质相互作用可以诱发蛋白质的交联、聚合,导致蛋白质共价结构变化、功能特性下降、风味恶化及营养损失,甚至产生有毒有害物质。研究表明,脂质氧化产物诱导的蛋白质氧化会造成色氨酸、精氨酸、酪氨酸等氨基酸含量减少;巯基含量增加、可溶性蛋白质的表面疏水性降低;α-螺旋、β-折叠增加;蛋白质聚集形成粒径较大的高分子聚集体。本文综述了脂质氧化产物-蛋白质相互作用及其对蛋白质结构和功能特性的影响,对于揭示脂质氧化产物与蛋白质的互作机制具有一定的指导意义。     

Antioxidant properties of aqueous and methanol soy extracts in minced trout muscle

Antioxidant capacity of freeze-dried aqueous and methanolic extracts of soy protein isolate (SPI) and soybean meal (SBM) were assessed by 2, 2′-Diphenyl-1-picrylhydrazyl (DPPH) assay and total phenolic content (TPC). Caffeic and chlorogenic acids in liquid aqueous extracts, and genistein and daidzein in liquid methanolic extracts were measured by HPLC. Freeze-dried extracts were applied (1000 and 4000 μg/g) to trout mince and TBARS formation monitored during 14 days refrigerated storage. SBM aqueous extracts-treated mince (4000 μg/g) had significantly lower (P < 0.05) TBARS than all other treatments. SPI methanolic extract had the highest TPC (100 mg chlorogenic acid equivalents/100 g extract) and genistein and daidzein contents. SPI aqueous extracts had the highest chlorogenic acid (280 μg/g) while SBM aqueous extracts had the highest caffeic acids content (484 μg/g). SPI had the best in vivo antioxidant profile. SBM extracts were most effective at retarding lipid oxidation in refrigerated trout mince.     

丙二醛氧化修饰对大豆蛋白热变性影响

以丙二醛代表脂质过氧化活性次生产物,采用圆二色谱、内源荧光光谱、粒径分析和差示扫描量热仪研究丙二醛氧化修饰对大豆蛋白热变性影响。结果发现,丙二醛氧化修饰使大豆蛋白热稳定性下降,相比大豆球蛋白,大豆β-伴球蛋白热稳定性下降更为显著。     

Effects of malondialdehyde-induced protein oxidation on the structural characteristics of rice protein

Malondialdehyde (MDA) was selected to represent a secondary by-product of lipid peroxidation during rice ageing. This study aimed to investigate the effects of MDA modification on the structural characteristics of rice protein. The results showed that as MDA concentration increased, rice protein carbonyl and disulphide groups increased, but sulphydryl content decreased. The blue shift of maximum fluorescence peak, the decrease of rice protein intrinsic fluorescence intensity and the reduction of surface hydrophobicity indicated the formation of protein aggregates caused by MDA oxidative modification. The results of molecular weight distribution and particle size distribution showed that MDA modification resulted in the formation of soluble protein aggregates, and the decrease of rice protein solubility indicated that insoluble protein aggregates were formed. Results of protein electrophoresis showed that MDA modification contributed to rice protein aggregation via non-disulphide covalent bonds. The results showed that rice protein gradually aggregated with increasing MDA concentration.     

Thermal behavior of emulsions manufactured with soy protein ingredients

The conformation, denaturation and aggregation behavior of proteins are important factors which dictate their ingredient functions and applications in formulated food products. The effect of variation in pre-treatment temperature (70–90 °C × 30 s), pH (6.4–7.5) and calcium supplementation (450 and 850 mg/L) on heat coagulation time (HCT at 140 °C) of model emulsions (3.6% (w/v) protein) stabilized with soy protein isolate (SPI) and soy protein hydrolysate (SPH) ingredients was determined. Generally, HCT of emulsions was not significantly affected by alteration of constituent pre-heating temperatures. Model emulsions displayed higher HCTs with increasing pH and lower levels of intrinsic ash content. At both supplementation levels, calcium addition led to decreased HCTs. Supplementation with chloride salts caused a greater decrease in HCT compared to supplementation with citrate salts. Furthermore, soy protein hydrolysis was associated with lower emulsion thermal stability. Results demonstrate that modification of ingredient and manufacturing parameters may be a useful approach for enhancing thermal stability properties of soy protein stabilized emulsions.     

Reactions of chlorogenic acid and quercetin with a soy protein isolate--influence on the in vivo food protein quality in rats

Plant phenolic compounds are known to interact with proteins producing changes in the food (e. g., biological value (BV), color, taste). Therefore, the in vivo relevance, especially, of covalent phenol-protein reactions on protein quality was studied in a rat bioassay. The rats were fed protein derivatives at a 10% protein level. Soy proteins were derivatized with chlorogenic acid and quercetin (derivatization levels: 0.056 and 0.28 mmol phenolic compound/gram protein). Analysis of nitrogen in diets, urine, and fecal samples as well as the distribution of amino acids were determined. Depending on the degree of derivatization, the rats fed with soy-protein derivatives showed an increased excretion of fecal and urinary nitrogen. As a result, true nitrogen digestibility, BV, and net protein utilization were adversely affected. Protein digestibility corrected amino acid score was decreased for lysine, tryptophan, and sulfur containing amino acids.     

Thermal stability of soy protein isolate and hydrolysate ingredients

The objective of this study was to characterize the effects of pH, protein concentration and calcium supplementation on thermal stability, at 140 °C, of soy protein isolate (SPI) and soy protein hydrolysate (SPH) ingredients. Increasing pH between 6.4 and 7.5 led to significantly (p < 0.05) higher mean heat coagulation times (HCTs) at 140 °C, for all soy protein ingredients at 1.8, and 3.6% (w/v) protein. Increasing protein concentration from 1.8 to 7.2% (w/v) led to shorter HCTs for protein dispersions. Calcium supplementation up to 850 mg/L, except in the case of supplementation of SPI 1 with calcium citrate (CaCit), decreased HCT for soy protein ingredient dispersions, at pH 6.4 – 7.5. No significant differences (p < 0.05) were found in mean HCT for dispersions supplemented with calcium chloride (CaCl₂) and those supplemented with CaCit at 450, 650 and 850 mg/L Ca²⁺, in the pH range 6.4–7.5.     

Antioxidative activity of some vegetable peels determined in vitro by inducing liver lipid peroxidation

Vegetable peels are normally thrown away as kitchen wastes. In order to reveal their possible biological value, an in vitro investigation was performed on the antioxidative potential of some peel extracts such as Luffa cylindrica (Lc), Raphanus sativus (Rs), Daucus carota (Dc), Pisum sativum (Ps) and Trichosanthes dioica (Td) in ferrous sulphate (FeSO₄), hydrogen peroxide (H₂O₂) and carbon tetrachloride (CCl₄) induced lipid peroxidation (LPO) in liver, the major target organ of any drug. Effects were compared with that of a known antioxidant, butylated hydroxy anisole (BHA). A dose dependent inhibition was observed in all three, FeSO₄, H₂O₂ and CCl₄ induced hepatic LPO. However, with respect to one test peel (Rs), the two lower doses (125 and 250 μg/ml) were found to be ineffective. We suggest that the test peels have the potential to inhibit the lipid peroxidation and the observed differential antiperoxidative effects of the test peels could be the result of their poly phenol and flavonoid contents.     

Preparation of isoflavones enriched soy protein isolate from defatted soy hypocotyls by supercritical CO2

Soy hypocotyls, the byproduct from soy protein industry, rich in isoflavones, was attempted to develop into a value-added functional food, soy protein isolate (SPI). Soy hypocotyls were defatted by supercritical CO₂ (SC-CO₂) under the following conditions as 35 MPa, 2 h, 45 °C with a constant flow rate of CO₂ at 5 l/min, fed from the bottom of the extraction vessel. Via extraction, the oil contents were lowered from 10.5 g/100 g in original soy hypocotyls to 0.2 g/100 g in defatted soy hypocotyls (DSH) with a Protein Dispersibility Index (PDI) value of 91.2 that contained about 2.18 g/100 g of isoflavones and 63.6 g/100 g of proteins. SPI was then extracted from DSH under the optimized conditions including: the pH of water solution at 6, the ratio of DSH to water solution as 1:10 and the extraction temperature at 30 °C with the extraction time of 30 min. The freeze-dried SPI contained 92.46 g/100 g of the proteins and 640 mg/100 g of isoflavones, where the three major β-glycosides, glycitin, daidzin and genistin accounted for about 83.7% of the total isoflavones. The yields of SPI and isoflavones were about 48% and 30% in terms of their respective total contents in DSH.     

Transglutaminase cross-linking to enhance elastic properties of soy protein hydrogels with intercalated montmorillonite nanoclay

To strengthen the network of bio-nanocomposite hydrogels, layered montmorillonite (MMT) nanoclay was intercalated by surface-coating with soy protein (SP) before mixing with 6% w/v SP for cross-linking by microbial transglutaminase (mTGase). Dynamic rheology was performed to study variables of NaCl and mTGase concentrations, with and without 1% w/v MMT. Without mTGase, the highest storage modulus (G′) was observed at 100 mM for samples without MMT, which was twice of the highest G′ for samples with MMT, at 200 mM NaCl. With mTGase, a shorter gelation time and a stronger hydrogel were observed at a higher enzyme level. Overall, the non-gelling 6% w/v SP dispersion was transformed to a hydrogel with G′ of 1099 Pa after addition of 100 mM NaCl and 1% SP-coated MMT and treatments by 6.25 U/g-protein mTGase for 2 h and heating/cooling steps. The integration of surface-coating and mTGase cross-linking is promising to improve properties of the nanocomposite system.     

Stabilization of acidic soy protein-based dispersions and emulsions by soy soluble polysaccharides

Soy soluble polysaccharides (SSPS) are shown to prevent destabilization of soy protein isolate (SPI) dispersions and SPI-based oil-in-water (O/W) emulsions under acidic conditions. Addition of SSPS above a critical concentration (0.25 wt%) increased the stability of 0.50 wt% SPI dispersions against aggregation and phase separation under conditions where SPI would normally precipitate (near its isoelectric point). Though SSPS neutralized SPI surface charge via electrostatic interaction, there was increased stability against aggregation due to steric repulsion. At acidic pH, addition of 1 wt% NaCl electrostatically screened protein–polysaccharide complexation which led to SPI precipitation and sedimentation. However, the order of salt addition had a significant impact on charge screening, with salt added before pH adjustment reducing SPI–SSPS complexation whereas it had less effect when added afterwards. Salt penetration efficacy diminished with decreasing pH. O/W emulsions (5 wt% oil) prepared with 0.50 wt% SPI destabilized at pH 4–5 due to protein aggregation, but addition of ≥0.25 wt% SSPS improved emulsion stability by inhibiting protein–protein interactions thus limiting increases in oil droplet diameter over time. Overall, both dispersion and emulsion stability greatly depended on pH, ionic strength and SSPS concentration. These results demonstrated that SSPS could effectively stabilize acidic SPI dispersions and that SPI–SSPS interactions may be used as a tool to improve the kinetic stability of SPI-based O/W emulsions.     

酶聚合结合超滤技术分离豆腐废水乳清蛋白的工艺研究

为改善豆腐乳清蛋白及低聚糖的回收率、提高膜通量,研究采用转谷氨酰胺酶在豆腐黄浆水自然条件下(温度50℃、pH值6.0)对乳清原液预处理,使蛋白质聚合,方便后续分离工艺选用截留分子量大的超滤膜分离大豆乳清蛋白。数据显示转谷氨酰胺酶Ⅰ可有效催化大豆乳清蛋白聚合,1%的酶添加量50℃反应30min即可催化95%以上的大豆乳清蛋白聚合,酶添加量3‰,聚合时间延长至5h。与对照组相比,乳清采用酶法预处理然后超滤分离,蛋白截留率及膜通量分别提高了2倍和1.3倍,而低聚糖的透过率没有明显影响。试验结果表明,相对于单纯的超滤工艺酶聚合预处理乳清然后超滤的分离效果是显著的。