Identification of aminoreductones as active components in Maillard reaction mixtures inducing nuclear NF‐κB translocation in macrophages

Coffee, a highly processed food, and Maillard mixtures are able to activate nuclear factor κB translocation in macrophages via generation of hydrogen peroxide. In this study, a substructure library was prepared and used to identify Maillard products that are responsible for this effect. Three different Maillard reaction products with aminoreductone substructure (C₆‐aminoreductone, C₄‐aminoreductone, and aminohexose reductone) strongly induce nuclear factor κB translocation in macrophages. The effect was almost completely blocked by co‐incubation with catalase, indicating that cellular activation was mediated by the ability of the test compounds to generate hydrogen peroxide. The cellular effect of a Maillard mixture, which was produced under conditions favoring aminoreductone formation, could be almost completely related to the presence of C₆‐aminoreductone.     

Effect of supercritical carbon dioxide treatment on the Maillard reaction in model food systems

The effect of supercritical carbon dioxide treatment on the development of the Maillard reaction in powdered model systems of lactose/ovine caseinmacropeptide and lactose/β-lactoglobulin at different pH values was studied. Supercritical carbon dioxide treatments in static conditions at 30 MPa and 50 °C for up to 5 h were applied to model systems. Control experiments at 50 °C were also performed. All assayed model systems treated with carbon dioxide under supercritical conditions showed lower extent of the Maillard reaction than control treated samples. Differences between supercritical carbon dioxide treated and control samples increased with pH. These results indicate that supercritical carbon dioxide treatment of food samples does not favour the Maillard reaction and thus can be applied in foods that may require special care to avoid excessive loss of available lysine.     

酪蛋白酸钠美拉德反应产物的制备及其乳化特性

利用小型超高温设备制备酪蛋白酸钠与葡萄糖、乳糖、低聚半乳糖、聚葡萄糖的美拉德反应产物,对比分析不同分子质量糖在不同热处理时间的美拉德反应进程及产物的乳化特性。结果表明,小分子质量的糖更易发生美拉德反应,褐变指数与反应程度呈正相关,乳化活性与接枝度均呈现先增大后减小的趋势,但两者并不成线性关系,酪蛋白酸钠与葡萄糖、乳糖、低聚半乳糖、聚葡萄糖乳化活性达到的最大值分别为0.63、0.51、0.55和0.48,其中130?℃热处理15?s的酪蛋白酸钠-葡萄糖溶液乳化活性最大,高于其他组,与水浴90?℃热处理90?min相当,并且乳化稳定性也呈现较高的水平,为123.88?min,将其用作乳化剂制备的DHA藻油乳状液稳定性动力学指数为1.5,显著小于其他组（P＜0.05）;由此可见,葡萄糖可作为美拉德反应的优良糖基配体制备新型高效的乳化剂,并且此方法可实现连续化生产,极大缩短了反应时间,提高了生产效率,对工业化生产具有指导意义。     

Effects of melamine on the Maillard reaction between lactose and phenylalanine

The effects of melamine on Maillard reaction were investigated in chemical model systems. The reaction products in the model systems with/without melamine were analysed by GC–MS and LC–MS/MS. Impact of melamine on Maillard browning in the above models was also examined by colourimetric methods. It was found that melamine can react directly with lactose and Strecker aldehydes formed in Maillard reactions to produce new adducts. In addition, the presence of melamine in chemical model systems also affects the formation of Maillard flavours and browning.     

甜菊苷对葡萄糖-氨基酸美拉德反应模型褐变强度及抗氧化活性的影响研究

作为低热高甜的新型糖源甜菊苷在食品工业中使用范围日益广泛,甜菊苷赋予食品甜感的同时也影响食品的加工品质,为探讨甜菊苷对美拉德反应的影响及在食品工业中的应用提供理论参考。通过建立葡萄糖与氨基酸之间的美拉德反应模型体系,考察甜菊苷添加量、反应温度、反应时长和初始pH条件对葡萄糖-甘氨酸、组氨酸、谷氨酸、半胱氨酸、苯丙氨酸美拉德反应褐变强度、抗自由基能力及总还原能力的影响。结果表明:在12mL 50mmol/L葡萄糖-氨基酸磷酸缓冲溶液反应体系中,甜菊苷添加量0.100g,加热温度为120℃、反应时间为90min、体系初始pH为8时,甜菊苷能够有效抑制褐变强度,抗氧化能力和反应历程因氨基酸类别而不同。为进一步研究甜菊苷在食品工业生产中与食品组分之间发生化学反应,以及在甜菊苷作用下美拉德反应机制奠定了初步理论基础。     

甜菊苷对葡萄糖-氨基酸美拉德反应模型褐变强度及抗氧化活性的影响研究

作为低热高甜的新型糖源甜菊苷在食品工业中使用范围日益广泛,甜菊苷赋予食品甜感的同时也影响食品的加工品质,为探讨甜菊苷对美拉德反应的影响及在食品工业中的应用提供理论参考。通过建立葡萄糖与氨基酸之间的美拉德反应模型体系,考察甜菊苷添加量、反应温度、反应时长和初始pH条件对葡萄糖-甘氨酸、组氨酸、谷氨酸、半胱氨酸、苯丙氨酸美拉德反应褐变强度、抗自由基能力及总还原能力的影响。结果表明:在12mL 50mmol/L葡萄糖-氨基酸磷酸缓冲溶液反应体系中,甜菊苷添加量0.100g,加热温度为120℃、反应时间为90min、体系初始pH为8时,甜菊苷能够有效抑制褐变强度,抗氧化能力和反应历程因氨基酸类别而不同。为进一步研究甜菊苷在食品工业生产中与食品组分之间发生化学反应,以及在甜菊苷作用下美拉德反应机制奠定了初步理论基础。     

Extrusion of soy protein with gelatin and sugars at low moisture content

Soy protein-based materials modified with gelatin, lactose and sucrose were prepared by extrusion at low moisture content. The effect of composition on the extrusion parameters was investigated and specific mechanical energy (SME) was measured as an indication of extrusion processability, thus providing good characterization of the extrusion process in order to make it highly energy efficient and cost effective. Water content was the most important factor on the extrusion parameters and product properties. The incorporation of gelatin increased SME and the product obtained at the extruder die was not continuous. However, when lactose was added, SME decreased and the color of the product changed due to Maillard reaction. This reaction could be analyzed by Fourier transformed infrared spectroscopy (FTIR) where the changes of the amide I and amide II bands reflected that hydroxyl groups in sugars and amino groups in soy protein isolate (SPI) were consumed during extrusion. These results are in good agreement with total soluble matter (TSM) values, which were lower for mixtures with lactose than sucrose due to a higher degree of Maillard reaction. Moreover, X-ray diffraction (XRD) and scanning electron microscopy (SEM) results also showed the influence of Maillard reaction, which lead to more ordered and compact structures, respectively.     

糖对谷氨酰胺转氨酶交联米渣蛋白美拉德反应产物结构与功能的影响

将不同相对分子质量的核糖、葡萄糖、乳糖、阿拉伯胶与谷氨酰胺转氨酶(transglutaminase,TG)交联的米渣蛋白(rice dreg protein,RDP)进行美拉德反应,并分析美拉德反应产物(Maillard reaction products,MRPs)的结构、功能特性。结果表明:不同相对分子质量的糖与TG交联RDP在美拉德反应过程中,随着反应的进行,接枝度、褐变度及体系的p H值有明显变化,其中核糖和葡萄糖的变化最为明显,其次是乳糖、阿拉伯胶。结构上,核糖最易与TG交联RDP中赖氨酸和精氨酸自由氨基发生反应;功能性质上,MRPs的溶解性和乳化特性较原TG交联RDP都得到了不同程度的改善,从强到弱依次是核糖、葡萄糖、乳糖、阿拉伯胶。所有样品的流变学特性都表现出假塑性流体所特有的剪切变稀现象,葡萄糖和核糖的MRPs表观黏度较原TG交联RDP大,乳糖和阿拉伯胶的MRPs的表观黏度则较原TG交联RDP小。     

Available lysine and fluorescence in heated milk proteins/dextrinomaltose or lactose solutions

In order to predict and compare the effects of dextrinomaltose and lactose on available lysine loss by the Maillard reaction, six model systems were prepared by mixing casein, laboratory whey protein or commercial whey protein with dextrinomaltose or lactose. The solutions were prepared at concentrations similar to those used in enteral and infant formula processing and were heated at 100, 120 or 140 °C for 0–30 min. The progress of the Maillard reaction in these model systems was followed by monitoring free fluorescence intermediary compounds. Model systems with lactose showed higher available lysine less than the model systems with dextrinomaltose; linear lysine losses were obtained between 0 and 20 min at 100 and 120 °C. At sterilization temperature and time (120 °C/10 min), lysine losses of milk proteins with dextrinomaltose as reducing sugar were 6.1% for casein, 4.1% for laboratory whey protein and 13.4% for commercial whey protein. Available lysine showed correlation with furosine in model systems prepared with lactose and casein or laboratory but not commercial whey protein at 100 and 120 °C. The initial fluorescence value obtained by mixing casein or laboratory whey protein with lactose or dextrinomaltose was low (between 3.8 and 5.7), whereas the value obtained when commercial whey proteins were used was close to 9. At 120 °C/10 min, there was only a small increase of fluorescence in casein and laboratory whey protein but a large increase in commercial whey protein (threefold the initial value). Fluorescence measurement is useful for finding the extent of the Maillard reaction in commercial whey protein (thermally damaged protein). An absolute value greater than 10 may indicate that products were prepared with thermally damaged proteins.     

Blocked Lysine in Dairy Products: Formation,Occurrence, Analysis,and Nutritional Implications

Lysine residues in milk proteins become “blocked” when they react with reducing sugars, particularly lactose, in the Maillard reaction. The blocked or glycated lysines reduce the biological availability of the lysine to metabolic processes and also hinder hydrolysis of the parent protein by digestive enzymes. Heating and storage of milk and milk products are the major promotants of the Maillard reaction and resulting chemical damage to the proteins. Several methods have been proposed to estimate the extent of this protein damage. Two major compounds, furosine, a product of acid hydrolysis of lactulosyl‐lysine, the 1st stable product of the Maillard reaction, and carboxymethyl‐lysine are used for assessing the early and advanced stages of the Maillard reaction, respectively. In addition, several methods are used for assessing the bioavailability of lysine in a protein; these include chemical, enzymic, and animal‐based methods. This review discusses the Maillard reaction and its significance in milk and dairy products, methods of assessing the extent of the reaction and of the bioavailability of lysine, and the nutritional significance of blocked lysines and associated Maillard reaction products in milk proteins.     

乳制品中美拉德反应机制及控制方法研究进展

乳制品经长时间储存或加热处理后容易发生美拉德反应,导致其营养价值下降,同时也会产生一些羟甲基呋喃,呋喃等对人体有害的物质,因此控制乳制品中美拉德反应对乳制品安全具有重要意义。乳制品中发生美拉德反应的主要成分为乳糖、乳清蛋白及酪蛋白等,且乳糖水解能够进一步促进美拉德反应发生。本文对乳制品中美拉德反应机制、美拉德反应产生的有害物质及控制措施进行了总结,并对乳制品美拉德反应的控制措施进行了重点阐述。本文旨在为乳制品体系中美拉德反应控制提供重要的科学依据。     

Immunochemical Properties of Proteins Glycosylated Through Maillard Reaction: β-Lactoglobulin-Lactose and Ovalbumin-Glucose Systems

β-Lactoglobulin and ovalbumin were glycosylated through Maillard reaction with glucose and lactose, respectively. The immunoreactivity between β-lactoglobulin-lactose and its specific antibody was inhibited strongly by free galactose and lactose, whereas free glucose did not at all inhibit the reaction between ovalbumin-glucose and its specific antibody. Furthermore, the antibody directed against β-lactoglobulin-lactose reacted also with other lactosylated proteins but the antibody to ovalbumin-glucose had no reactivity with the other proteins glycosylated with glucose, indicating that lactose attached to carrier proteins through Maillard reaction could be a hapten-like antigen but glucose could not.     

110℃下木糖-半胱氨酸美拉德反应模型风味物质及杂环胺的形成

为进一步实现美拉德反应的定向控制,试验采用牛肉汤提取物为原液,以OD值和感官评价为评定指标,通过析因试验设计6组不同质量浓度下木糖和半胱氨酸的美拉德反应模型,并采用固相微萃取-气相色谱-质谱联用技术(SPME-GC-MS)、超高效液相色谱-质谱联用技术(UPLC-MS)测定其挥发性风味物质和杂环胺的种类和含量。结果表明,在pH7,反应温度110 ℃,反应时间70 min的条件下,木糖和半胱氨酸浓度均为0.04 g/mL时,获得的美拉德反应模型评分最高,此组合中共鉴定出11类、52种挥发性风味物质,主要包括噻唑类(24.910%)、酮类(20.220%)、吡嗪(19.531%)和呋喃(14.606%)等。美拉德反应模型产生杂环胺2.660 ng/mL。本试验在较低温度下获得一种具备理想风味且杂环胺含量较低的美拉德反应模型产物,为美拉德反应的定向控制及其在肉制品中的应用提供理论参考。     

Effect of heating on Maillard reactions in milk

Heated milk is subject to the Maillard reaction; lactose and lysine residues in milk proteins (mainly casein) are the reactants. An overview is given of the early, advanced and final stages of the Maillard reaction as it occurs in milk. The early Maillard reaction is confined to the formation of the protein-bound Amadori product lactulosyllysine. Breakdown of the Amadori product leads to formation of all kinds of advanced Maillard reaction products such as lysylpyrraline, pentosidine, hydroxymethylfurfural, (iso)maltol, furfurals and formic acid. The content of these compounds in heated milk is, however, very low (with the exception of formic acid), and does not correspond to the breakdown of Amadori product in quantitative terms. The final stage, in which melanoidins (brown pigments) are formed and protein polymerization occurs, is largely unknown from a chemical point of view, let alone quantitatively. The conclusion can only be that not all important compounds are yet identified. Some experimental data for heated milk are given to illustrate the various stages of the Maillard reaction in heated milk. A kinetic analysis of the Maillard reaction is difficult because it is such a complicated reaction with many parallel and consecutive steps; in addition, one of the reactants, lactose, is also subject to another reaction, namely isomerization followed by degradation. The kinetics can be tackled by kinetic, multiresponse modelling, and this approach is illustrated. It appears that the temperature dependence of the (early) Maillard reaction is lower than for the simultaneously occurring isomerization reactions of lactose. The use of several components formed in the Maillard reaction to evaluate the heat intensity given to milk is discussed.     

改性乳清蛋白体外抗氧化特性

以木糖、葡萄糖、果糖、乳糖、麦芽糖、蔗糖和乳清蛋白为美拉德反应原料,按照一定比例进行湿热糖基化反应。结果表明:在湿热糖基化反应后,6种糖类与乳清蛋白复合物在420 nm和294 nm处的吸光值差异显著(P<0.05);除乳清蛋白-蔗糖复合物外,其他5种乳清蛋白糖基复合物的游离氨基酸含量均显著降低(P<0.05)。除了乳清蛋白-蔗糖复合物外,其他5种乳清蛋白糖基化复合物随浓度的增加,还原能力和DPPH自由基清除能力都有所提高,其中乳清蛋白-木糖复合物的抗氧化能力提高最大,而且SDS-PAGE显示乳清蛋白-木糖复合物反应后分子质量明显增大;傅里叶红外光谱(FTIR)结果显示,乳清蛋白-木糖和乳清蛋白-葡萄糖复合物中蛋白质的酰胺I和II明显减少。     

Characterization and improvement of rheological properties of sodium caseinate glycated with galactose, lactose and dextran

The aim of this work was to investigate the effect of non-enzymatic glycosylation with galactose, lactose, and 10 kDa dextran on the rheological properties of sodium caseinate. To promote the formation of covalent complexes, the reaction was done in solid state (a_w = 0.67), pH 7.0 (0.1 M sodium phosphate buffer), and temperature set at 50 and 60 °C. The progress of Maillard reaction was indirectly traced by measuring the formation of the Amadori compound, through furosine (2-furoylmethyl-lysine) analysis, and brown polymers, and the resulting glycoconjugates were characterized by LC/ESI-MS and SEC. Results showed a higher reactivity of galactose than lactose and dextran to form the glycoconjugates, due to its smaller molecular weight. Glycation with galactose and lactose increased the viscosity of caseinate and also altered its flow characteristics from Newtonian to shear-thinning. Oscillatory testing showed a higher elastic modulus (G′) in glycoconjugates when compared to non-glycated caseinate, especially with galactose, where a gel-like behaviour was observed after long incubation times. Glycation with dextran did not produce substantial improvements in the rheological properties of caseinate, probably due to the limited extent of the reaction. Our results show that by controlling the rate and extent of the Maillard reaction is a technologically feasible operation to improve the viscosity and gelling properties of sodium caseinate-based ingredients.     

Fluorescence associated with Maillard reaction in milk and milk-resembling systems

The progress of the Maillard reaction in milk and milk-resembling systems during heating was followed by monitoring free fluorescent intermediary compounds. Fluorescence intensity was examined over a wide temperature/time range (90–140 °C/0.5–30 min). Different kinetic behaviours were found in the presence or absence of amino groups in the system. Traces of fluorescence were detected when the lactose system without proteins was heated; the isomerization and degradation reactions of lactose could also generate fluorescent compounds. Fluorescence accumulation in the lactose system (without proteins) proceeded according to first-order reaction kinetics, whereas systems with lactose and casein, lactose and whey protein and milk fitted zero-order reaction kinetics. Apparent activation energies calculated were 61.3, 78.6, 59.6 and 83.9 kJ mol⁻¹ for lactose system, lactose/casein system, lactose/whey protein system and milk, respectively. Moreover, levels of intermediary fluorescent compounds in Spanish commercial milk (pasteurized, UHT-treated and in-bottle sterilized milk) have been studied.     

Effect of drying methods on the reactivity of chitosan towards Maillard reaction

The effect of drying methods on chitosan reactivity towards Maillard reaction during storage of dried chitosan–lactose systems was investigated. Two different structural forms of chitosan, scaffold and microspheres, were prepared. Then they were dried by lyophilization or using supercritical fluid technology (SF-CO₂) and stored with lactose under controlled temperature conditions (60 °C) and water activity (a_w = 0.65). The drying method produced slight modifications on the chitosan structure especially in samples submitted to SF-CO₂ treatment. Differences were more evident in the reactivity of chitosan–lactose systems during storage. Maillard reaction development was assessed by quantification of a new compound which originated from the acid hydrolysis of Amadori compounds (NFMD) resulting from chitosan–lactose interactions, and not detected in the acid hydrolysates of freshly prepared chitosan-lactose systems. Maillard reaction was influenced by different factors such as the manner of lactose addition, structure of chitosan (microspheres and scaffolds) and drying method. Lyophilized chitosan in the form of microspheres with lactose, presented the highest values of NFMD (428.45 mg/100 g of sample). The determination of this compound could be used as an index of the progress of Maillard reaction.     

氨基酸对浓香葵花籽油美拉德反应风味的贡献

通过研究比较脱脂葵花籽粕烘烤前后的氨基酸组成及风味变化,建立了以浸出一级葵花籽油为基质的氨基酸与葡萄糖美拉德反应风味模拟模型,比较分析了12种氨基酸与葡萄糖美拉德反应风味模拟模型体系中形成的挥发性化合物与浓香葵花籽油风味化合物的异同。结果表明:赖氨酸、精氨酸、组氨酸对浓香葵花籽油美拉德反应风味的贡献最大,是浓香葵花籽油美拉德反应风味形成的主要前体物质。赖氨酸、精氨酸、组氨酸3组模型体系中美拉德反应挥发性化合物与浓香葵花籽油风味化合物相同的共有12种,其中吡嗪类化合物8种、醛类化合物2种、醇类化合物1种、呋喃类化合物1种。     

NMR和GC-MS研究加热对奶油主要成分及香气化合物的影响

为深入了解加热过程中奶油主要成分和香气成分的变化机理，对加热前后奶油进行分析。利用核磁共振（nuclear magnetic resonance，NMR）技术分别对奶油中的极性组分和非极性组分进行定量分析，同时利用气相色谱-质谱（gas chromatography-mass spectrometry，GC-MS）联用技术对加热前后奶油中的关键香气化合物进行鉴定。NMR分析表明，奶油中的极性组分主要为乳糖以及少量的游离氨基酸、游离脂肪酸和有机酸，非极性组分主要为甘油三酯以及少量的双甘酯和单甘酯。经加热后，极性组分含量大量减少，主要为乳糖和游离氨基酸，而非极性组分含量减少相对较小；GC-MS分析表明，短链脂肪酸和醛类含量增加相对较少，甲基酮、内酯和美拉德反应产物含量增加显著，其中甲硫基丙醛、麦芽酚和呋喃酮仅在加热后奶油中检出。结果表明加热过程中美拉德反应产物的形成和脂质来源化合物的增加分别造成了奶油中极性组分和脂肪酸的减少。该研究结果为奶油风味及产品研究提供了思路。