不同加工方式降低大黄鱼鱼卵过敏原及其 消化产物免疫反应性的比较研究

目的 本文以大黄鱼鱼卵为原料, 分析比较不同加工方式对大黄鱼鱼卵过敏原及其消化产物免疫反应性的影响。方法 采用不同加工方式(美拉德反应, 紫外照射, 超声联合加热, 高温高压)处理卵黄蛋白,制备粗提物并进行模拟胃肠液二相消化实验, 利用免疫印迹及抑制性ELISA方法对粗提物及消化产物中过敏原的IgG/IgE结合活性进行分析。结果 分析不同加工方式处理后卵黄蛋白中过敏原的IgG结合活性, 结果发现当抗体稀释倍数为4×10₄时, 检测不到美拉德反应产物与抗体发生特异性结合, 而经后三种物理加工方式处理后的粗提物均与抗体有不同程度的免疫反应。此外, 分析加工处理及其消化产物中过敏原与患者血清的IgE结合活性, 结果显示经不同加工方式处理后, 仅有美拉德反应产物的抑制率低于50%; 经高温高压处理的粗提物抑制率为65%, 但消化后其抑制率降至20%以下。结论 在4种不同的加工方式中, 美拉德反应能有效地降低大黄鱼鱼卵中过敏原的IgG/IgE结合活性, 超声联合加热及高温高压处理明显减弱过敏原消化产物的IgE结合活性, 紫外照射对过敏原的影响不明显。     

体外人胃肠模拟系统在食物消化行为研究中的应用进展

人体胃肠道在食物消化和营养物质的吸收过程中具有非常重要且关键的作用,在对人体胃肠道功能进行研究过程中常存在伦理问题,而体外人胃肠模拟系统可有效解决该问题,对促进胃肠道功能的研究和食物消化行为的研究也起着非常重要的作用。该文对体外人胃肠模拟系统的研究现状、研究局限性等进行了综述,并对体外人胃肠模拟系统的应用研究进行展望,提出一些发展建议和意见,以期促进体外人胃肠模拟系统在食物消化行为研究方面的工作。     

Antioxidant activity of amaranth protein or their hydrolysates under simulated gastrointestinal digestion

Amaranth proteins were subjected to a simulated gastrointestinal digestion to evaluate the antioxidant activity of the products. A protein isolate (I) was first hydrolyzed with pepsin (Pe) (pH 2, 37 °C) and then with pancreatin (Pa) (pH 6, 37 °C). Different hydrolysis conditions were assayed and control reactions (without enzymes) were performed. Hydrolysis degree (HD) determined by TNBS method ranged from 13 to 37%. Soluble fractions in 35 mmol/L phosphate buffer, pH = 7.8 were obtained from freeze-dried samples, and antioxidant activity was evaluated by the ABTS⁺·scavenging and the ORAC assays. Antioxidant activity increased significantly (p < 0.05) after simulated gastrointestinal digestion. According to the results, digestion conditions (Pe/protein: 1:10, 60 min; and Pa/protein: 1:10, 60 min) were selected and applied to an amaranth protein alcalase-hydrolysate (H) (HD = 29.2 ± 1.3). After pepsin and pancreatin action (Hpepa), HD was 42.0 ± 2.6, slightly higher than that of the digested isolate (Ipepa) (36.9 ± 0.5). The corresponding soluble fractions exhibited different electrophoretic profiles (tricine-SDS-PAGE) and gel filtration chromatograms, evidencing the presence of different molecular species. Previous hydrolysis with alcalase did not improve the antioxidant activity after simulated gastrointestinal digestion according to the methodologies assayed. Both the protein isolate and the alcalase-hydrolysate showed a potential capacity to scavenge free radicals after gastrointestinal digestion, appearing as promising ingredients to formulate functional foods with antioxidant activity.     

马鹿茸水溶性蛋白的提取与体外模拟消化研究

以鲜马鹿茸为原料,对比六种蛋白提取方法,依据提取蛋白量及SDS-PAGE电泳图谱确定鹿茸水溶性蛋白有效提取方法;模拟胃肠环境对提取的蛋白进行体外模拟消化研究,测定其体外胃肠消化稳定性及消化过程中游离氨基含量变化。结果表明:组成为100mmol/L Tris、6mol/L盐酸胍、0.02mol/L EDTA-2Na和1%胃蛋白酶抑制剂的提取液提取蛋白量最高,电泳条带多且清晰;蛋白在体外模拟胃、肠环境下消化5min后即迅速被分解,消化60min后即被分解为20ku以下的小分子量蛋白或多肽,消化2.5h后分子量主要分布在7823u及以下范围,在肠和胃肠连续消化过程中游离氨基含量随时间的延长而显著增加,这表明鹿茸水溶性蛋白较易被分解;模拟肠消化2.5h后游离氨基含量增加了一倍,显著大于胃消化中的增加量（p<0.01）,这暗示着鹿茸水溶性蛋白更有利于肠消化。      

Comparison of antioxidant activities of bovine whey proteins before and after simulated gastrointestinal digestion

Oxidative stress caused by free radicals has been implicated in several human disorders. Dietary antioxidants can help the body to counteract those reactive species and reduce oxidative stress. Antioxidant activity is one of the multiple health-promoting attributes assigned to bovine whey products. The present study investigated whether this activity was retained during upper gut transit using a static simulated in vitro gastrointestinal digestion (SGID) model. The capacity to scavenge free radicals and reduce ferric ion of whey protein isolate (WPI), individual whey proteins, and hydrolysates pre- and post-SGID were measured and compared using various antioxidant assays. In addition, the free AA released from individual protein fractions in physiological gut conditions were characterized. Our results indicated that the antioxidant activity of WPI after exposure to the harsh conditions of the upper gut significantly increased compared with intact WPI. From an antioxidant bioactivity viewpoint, this exposure negates the need for prior hydrolysis of WPI. The whey protein α-lactalbumin showed the highest antioxidant properties post-SGID (oxygen radical absorbance capacity = 1,825.94 ± 50.21 μmol of Trolox equivalents/g of powder) of the 4 major whey proteins tested with the release of the highest amount of the antioxidant AA tryptophan, 6.955 μmol of tryptophan/g of protein. Therefore, α-lactalbumin should be the preferred whey protein in food formulations to boost antioxidant defenses.     

The effect of various heat treatments on the antioxidant capacity of milk before and after simulated gastrointestinal digestion

Various reactions take place between compounds of milk during heat treatment including denaturation and aggregation of whey proteins, and formation of new complexes. In this study, the antioxidant capacities of raw, pasteurised and sterilised milk were determined both before and after an enzymatic in vitro digestion process, using a radical cation decolorisation assay. The mean values of raw, pasteurised and sterilised milk before simulated digestion were 4.02, 4.47 and 4.18 mM trolox/g, respectively, and these did not differ significantly. The antioxidant capacity of all milk types increased significantly at the end of the simulated gastrointestinal digestion, being 11.13, 12.33, 11.88 mM trolox/g for raw, pasteurised and sterilised milk, respectively.     

Effect of thermal processing and digestive protease on the antioxidant capacity of fruit juice–milk beverage model systems under simulated gastrointestinal digestion

The effects of thermal processing and digestive protease on the antioxidant capacity of fruit juice–milk beverage (FJMB) models containing whey protein (WP) and chlorogenic acid or catechin (CAT) under in vitro digestion were investigated. After gastric digestion, the FJMB showed a significant (P < 0.05) increase in ABTS, but little change in ferric reducing antioxidant power (FRAP). Intestinal digestion decreased the FRAP and increased ABTS except in the model containing 0.1% CAT. There were different effects (none, masking, synergistic effect) in the antioxidant activity of the WP and phenolics under digestion. Pasteurisation (63 °C/30 min) had no significant effect on the FJMB's antioxidant capacity, while sterilisation (121 °C/10 min) significantly (P < 0.05) increased, decreased or had no significant effect on ABTS and FRAP under digestion, depending on the model. Pepsin and pancreatin significantly (P < 0.05) decreased the fruit phenolics' ABTS. Ferric reducing antioxidant power was not significantly influenced by pepsin during gastric digestion, but was significantly (P < 0.05) decreased by pancreatin during intestinal digestion. These results would be helpful to improve the efficacy of fruit phenolics in FJMB subjected to thermal processing and to maintain the health benefits of this kind of functional products in the food industry.     

鱼肉蛋白肽在模拟胃肠消化吸收过程中的抗氧化活性和生物利用度

以鳕鱼鱼肉蛋白肽为研究对象,构建体外胃肠消化模型和Caco-2细胞吸收模型,模拟连续的胃肠消化、吸收过程,测定抗氧化活性和生物利用度。结果显示:在模拟胃肠消化过程中,鱼肉蛋白肽的水溶性维生素E抗氧化能力（TEAC）变化不显著（p>0.05）,DPPH自由基清除率在胃消化阶段下降而在肠消化阶段升高（p<0.05）,但低于消化前的水平（p<0.05）。在模拟转运2 h过程中,鱼肉蛋白肽吸收产物的肽氮含量逐渐增加,TEAC和氧自由基抗氧化能力（ORAC）显著升高（p<0.05）。鱼肉蛋白肽的生物利用度显著高于鱼肉（9.1%）,且分子量最小的蛋白肽的生物利用度最高（46.2%）。鱼肉蛋白肽经胃肠消化吸收后,具有一定的抗氧化活性且生物利用度较高,为鳕鱼蛋白肽的开发利用提供了理论依据。      

体外模拟消化条件下岩藻黄素产品抗氧化活性的变化

目的 评价岩藻黄素产品在消化系统中的稳定性及变化规律。方法 本文考察了在体外消化模拟状态下岩藻黄素产品中岩藻黄素含量变化, 并采用测定其对DPPH自由基清除能力、铁离子还原能力(ferric ion reducing antioxidant power, FRAP)与氧自由基吸收能力 (oxygen radical absorbance capacity, ORAC)这3种抗氧化指标来评价岩藻黄素产品的抗氧化能力变化。结果 随着人工模拟胃肠消化过程时间的增加, 岩藻黄素产品中岩藻黄素的含量呈下降趋势, 但岩藻黄素微囊粉较岩藻黄素油悬液对岩藻黄素表现出良好的包埋与保护作用。在胃肠消化过程中, 岩藻黄素微囊粉与油悬液对DPPH自由基清除能力、FRAP铁离子还原能力与ORAC氧自由基吸收能力等抗氧化活性指标均有所降低, 其中岩藻黄素微囊粉较油悬液稳定, 表现出较好的抗氧化活性。结论 岩藻黄素微囊粉产品比油悬液产品在消化体系中具有更好的稳定性。     

Protein quality and antioxidant properties of soymilk derived from black soybean after in vitro simulated gastrointestinal digestion

Protein quality and antioxidant properties of soymilk derived from black soybean (eight varieties) in China were analysed following in vitro simulated gastrointestinal digestion (including dialysis). Soymilk from black soybean possessed high okara weight but low yield, protein content and sensory scores. The in vitro digestibility of protein in all black soymilk samples was higher than 60%, and the Shenmu black soybean exhibited the highest digestibility. Non-digested milk from the black soybean exhibited significantly high total phenolic content (TPC) (127.15–173.04 mg/100 mL), ferric-reducing antioxidant power (FRAP) (272.18–366.27 μmol L⁻¹) and DPPH free radical-scavenging activity (61.20–83.81%). These parameters were significantly lower in the non-digested soymilk than those in soymilk after gastric digestion but higher than those of soymilk in the dialysed fraction. Gastric digestion significantly increased bioactive compound levels released from black soymilk, and the bioaccessibility of phenolic compounds was 24.37–36.05%. Hence, black soymilk was sufficiently available for human absorption.     

Angiotensin converting enzyme-inhibitory activity of peptides isolated from Manchego cheese. Stability under simulated gastrointestinal digestion

In this study, several peptides, which had previously been identified in active HPLC fractions from Manchego cheese, were synthesised and their angiotensin converting enzyme (ACE)-inhibitory activities were measured. From 11 peptides, which were selected based on their structures, only two, VRYL and KKYNVPQL, showed considerable ACE-inhibitory activity with IC₅₀ values of 24.1 and 77.1 μ , respectively. Subsequently, the impact of the gastrointestinal digestion on ACE-inhibitory activity was evaluated. Some of the peptides selected were resistant to the incubation with pepsin followed by hydrolysis with a pancreatic extract. The ACE-inhibitory activity after simulated digestion did not change drastically except for peptide _s2-CN f(195-204) (TQPKTNAIPY) that exhibited an activity 6 times greater after simulated digestion. In contrast, after simulated digestion, the activities of peptides VRYL and KKYNVPQL decreased. The peptides not hydrolysed by gastrointestinal enzymes and peptide VRYL, which was only partly hydrolysed, were incubated with ACE and were found to be true inhibitors of the enzyme and to have a competitive inhibition pattern.     

Effect of food matrix and processing on release of almond protein during simulated digestion

The aims of the present work were to assess digestibility of almond protein in the upper gastrointestinal tract, evaluate the effects of food matrix on protein release and assess the persistence of immunoreactive polypeptides generated during simulated digestion. Prunin, the most abundant protein in almond flour, was sensitive to pepsin, with complete digestion after 20 min in the gastric phase. Addition of the surfactant phosphatidylcholine did not affect the rate and kinetic of digestion, as observed by SDS-PAGE analysis and HPLC, in the stomach and the small intestine of either natural or blanched almond flour. However, incorporation of almond flour into a food matrix, such as chocolate mousse and Victorian sponge cake, decreased the rate of almond protein degradation by pepsin and immunoreactivity of almond polypeptides detected by dot blots and sandwich ELISA retained better. Most of the almond protein identified by in-gel tryptic digestion and MALDI-TOF analysis corresponded to prunin, with pI values of 5–7.     

紫苏油微胶囊的性质与体外模拟消化研究

为提高紫苏油在人体胃肠道内的有效消化吸收能力,以明胶和海藻酸钠为壁材,采用复凝聚法制备紫苏油微胶囊,并对微胶囊的理化特性、抗潮性、耐热性、模拟胃肠道条件下的消化吸收能力以及微胶囊化前后紫苏油的脂肪酸成分进行分析。结果表明：紫苏油微胶囊水分含量为3.91%,堆积密度为0.31 g/mL,休止角为44.47°,包埋率为87.747%。微胶囊产品应尽量置于阴凉干燥的环境中保存,在食品加工中,加工温度应控制200℃以下,且尽可能缩短加工时间。在模拟消化吸收过程中,胃液内累积释放率为 13.25%;肠液内累积释放率33.02%;释放机理符合Ritger-Peppas模型,释放机制为非Fick 扩散过程。微胶囊化前后紫苏油的脂肪酸含量基本无变化。     

Identification of casein phosphopeptides after simulated gastrointestinal digestion by tandem mass spectrometry

The bovine casein was subjected to in vitro gastrointestinal digestion simulating physiological conditions. Out of a total of 19 different casein phosphopeptides (CPPs), 10 had been found with the cluster sequence S(P)S(P)S(P)EE representing the binding site for minerals, which were identified by on-line reversed-phase high performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (RP-HPLC-ESI-MS/MS). Twelve CPPs formed in this study are reported for the first time and four of them (α_s1-CN(57-90)5P, α_s1-CN(56-90)5P, α_s1-CN(55-76)5P, β-CN(1-52)5P) contained the cluster sequence S(P)S(P)S(P)EE. The potential mineral chelating properties of CPPs identified in relation to their amino acid sequences and the presence of the phosphorylated cluster is discussed.     

The simulated in vitro infant gastrointestinal digestion of droplets covered with milk fat globule membrane polar lipids concentrate

Milk fat globule membrane polar lipids (MPL) are increasingly used as the surface-active components for emulsions in many infant food products. However, the precise effect of the emulsifier MPL on the digestion of lipids during gastrointestinal digestion has not been elucidated. This study investigated the lipid digestion of droplets covered with MPL with different sizes in a simulated in vitro infant gastrointestinal digestion assay. The well-used surface-active component casein was used as a control. Four types of emulsions were formulated: small and large droplets covered with MPL concentrate (MPL-S and MPL-L, with volumetric means of 0.35 ± 0.01 and 4.04 ± 0.01 μm, respectively), and small and large droplets covered with casein (CN-S and CN-L, with volumetric means of 0.44 ± 0.01 and 4.09 ± 0.03 μm, respectively). The emulsions were subjected to in vitro gastrointestinal digestion using a semidynamic model mimicking infant digestion. Through the determination of particle size evolution, zeta-potential, and microstructure of emulsions, the lipid droplets covered with MPL were found to be more stable than that of the CN-S and CN-L during gastrointestinal digestion. Moreover, although CN-S and CN-L showed a higher initial lipolysis rate at the beginning of gastric digestion, droplets covered by MPL exhibited a significantly higher amount of free fatty acid release during later digestion. The amount of free fatty acid release of the emulsions in both gastric and intestinal digestion could be generally classified as MPL-S ≥ MPL-L > CN-S > CN-L. Our study highlights the crucial role of MPL in the efficient digestion of emulsions and brings new insight for the design of infant food products.     

Symposium review: Fat globules in milk and their structural modifications during gastrointestinal digestion

The fat globules in milk are unique oil droplets that are stabilized by a specific and structurally complex membrane, the milk fat globule membrane (MFGM). In the last decade, excellent progress has been made on studying the structure of the milk fat globules and the MFGM and how common processing treatments affect these structures to deliver dairy products with improved functional properties. Although the digestion of milk fat to deliver energy and lipid-soluble nutrients is essential for survival of the neonate, there is little understanding of the complex processes involved. The structural alterations to fat globules during gastrointestinal processing affect the way in which milk fat is digested, absorbed, and metabolized. The packaging of these globules within the MFGM or in other forms may affect the bioaccessibility of raw or processed milk fat globules; in turn, this may affect access of the gastrointestinal enzymes to the globules and, therefore, may influence the rate and extent of lipid digestion. This review focuses on recent advances in understanding milk fat globules during gastrointestinal digestion, including the effects of processing on their bioavailability and the kinetics of lipid digestion. Possible effects of the dairy matrix on lipid digestion and physiological responses are briefly described.     

体外模拟消化对酸枣仁蛋白酶解产物抗氧化活性的影响

目的:以具有良好抗氧化活性的酸枣仁蛋白酶解产物(SZPHs)为研究对象,探究体外模拟胃肠消化对其氨基酸组成及抗氧化活性的影响.方法:通过体外模拟胃肠消化,得到酸枣仁蛋白酶解产物消化物(SZPHs-GD),并分析SZPHs消化前后的氨基酸组成以及体外抗氧化活性.结果:体外模拟胃肠消化后,SZPHs-GD中疏水性氨基酸含量...