基于葡萄糖双子表面活性剂和卵白蛋白的生物活性物质载体制备和特性研究

发布时间：2018-02-20 04:41

  本文关键词： 葡萄糖双子表面活性剂 卵白蛋白 胶束 囊泡 纳米凝胶 姜黄素 儿茶素酯 鱼油　出处：《浙江大学》2017年博士论文　论文类型：学位论文

【摘要】：本文以葡萄糖双子表面活性剂和卵白蛋白为主要壁材,构建了胶束、囊泡、聚电解质包覆囊泡、纳米颗粒、纳米凝胶和微乳液等一系列生物活性物质载运体系。在此基础上,研究了它们对姜黄素、儿茶素酯和鱼油等生物活性物质的载运能力以及稳定性、功能性调控作用,并探讨了相关机理,主要结论如下:(1)制备了葡萄糖双子表面活性剂/季铵盐表面活性剂混合胶束,并考察了混合胶束包埋对负三价姜黄素(Cur~(3-))的稳定性提高作用。结果表明,由于混合胶束分子之间的静电斥力比季铵盐表面活性剂胶束分子之间的静电斥力弱,混合胶束的栅栏层更加致密且疏水性更强。另外,与季铵盐表面活性剂胶束相比,混合胶束与Cur~(3-)的静电相互作用较弱但疏水相互作用更强,使得Cur~(3-)更容易进入混合胶束栅栏层内部。因此,混合胶束可以更加有效地将Cur~(3-)与外界环境隔离并抑制其降解。(2)研究了以十二醇葡萄糖双子表面活性剂(Gemini-C12)为主要壁材的类脂质囊泡对儿茶素酯的载运能力和抗氧化活性提高作用。结果表明,儿茶素酯在囊泡双分子层中采取了垂直定位的方式,因此它在囊泡中的有效载量较高。在一定的载量范围内,儿茶素酯可以提高囊泡双分子层的有序性和相变温度。当超过这个范围时,儿茶素酯荷载增加了双层膜的流动性并且降低了其热稳定性。囊泡包封儿茶素酯在脂质体模型中的抗脂质氧化的能力高于游离儿茶素,这可能与儿茶素酯倾向于定位在磷脂双分子层中有关。(3)通过静电修饰的方法构建了羧甲基纤维素钠(CMC-Na)包覆囊泡,并研究了 CMC-Na包覆对囊泡稳定性的提高作用。结果表明,CMC-Na与囊泡的结合是一个自发的吸热过程,由熵变驱动。通过控制CMC-Na浓度可以获得尺寸小、性质稳定的包覆囊泡,并且实验获得的CMC-Na最佳浓度与经验模型预测的CMC-Na饱和浓度基本吻合。CMC-Na修饰提高了囊泡的pH稳定性、热稳定性和盐稳定性,并提高了芯材儿茶素酯在热处理条件下的保留率。(4)研究了 Gemini-C12与热变性卵白蛋白(OVA)的相互作用并探讨了Gemini-C12浓度对Gemini-C12-OVA复合物包覆小麦醇溶蛋白(gliadin)纳米颗粒性质的调控机理。结果表明,当OVA-Gemini-C12混合溶液中Gemini-C12浓度低于临界胶束浓度(CMC)时,游离Gemini-C12倾向于吸附在颗粒-水界面。此时,Gemini-C12可以提高反溶剂过程中的成核速率,从而减小gliadin纳米颗粒的尺寸;另一方面,它可以填充颗粒表面OVA吸附层留下的空隙,从而提高了纳米颗粒的贮藏稳定性和再分散性。当Gemini-C12浓度高于CMC时,表面活性剂分子倾向于定位在胶束中而不是吸附在固-液界面上,从而增加了 gliadin纳米颗粒的尺寸并降低了其贮藏稳定性和再分散性。(5)结合美拉德反应和等电点加热凝胶法制备了卵白蛋白-葡聚糖纳米凝胶,并评价了纳米凝胶包封对姜黄素生物利用率的提高作用。卵白蛋白-葡聚糖纳米凝胶主要呈球形结构,其pH稳定性、贮藏稳定性和再分散性均较好。通过pH驱动法实现了纳米凝胶对姜黄素的荷载,纳米凝胶在包封姜黄素后理化性质无明显变化。卵白蛋白-葡聚糖纳米凝胶和卵白蛋白纳米颗粒包封均可以提高姜黄素在模拟消化中的转化率(transformation)和生物可给率(bioaccessibility)。纳米凝胶包封姜黄素比纳米颗粒包封姜黄素的转化率更高,而两者之间的生物可给率无显著差异,这可能和它们在模拟消化后相似的理化性质有关。(6)通过自由基枝接法制备了卵白蛋白与儿茶素的共价结合物,并研究了卵白蛋白和三种共价结合物对鱼油乳液的稳定作用。由于酚羟基的引入,卵白蛋白的自由基清除能力得到了提高。儿茶素枝接改变了卵白蛋白的二级结构,并提高了其表面疏水性和三级构象柔顺性。与卵白蛋白鱼油乳液相比,卵白蛋白-儿茶素共价结合物鱼油乳液的脂滴更小、贮藏稳定性更好、粘度更低并且氧化稳定性更高。总体而言,鱼油乳液在pH 3.5条件下的氧化稳定性比pH 7.0条件下高,这可能主要与pH3.5条件下带正电的脂滴对过渡金属的排斥作用有关。
[Abstract]:In this paper, glucose Gemini surfactants and ovalbumin as main wall material, construction of the micelles, vesicles, polyelectrolyte coated vesicles, nano particles, nano and micro emulsion gel and a series of bioactive substances carrying system. On this basis, the research on carrying capacity of curcumin, catechin esters and other fish biology active substance and stability, functional regulation, the relevant mechanism and discussion, the main conclusions are as follows: (1) glucose Gemini Surfactant / quaternary ammonium surfactant mixed micelles were prepared, and the effects of the mixed micelle entrapped curcumin on negative trivalent (Cur~ (3-)) improved the stability. Results show that due to the electrostatic repulsion between molecules in the mixed micelles than the electrostatic repulsion between quaternary ammonium surfactant molecules is weak, the palisade layer of the mixed micelle is more dense and more hydrophobic. In addition, with quaternary ammonium salts Compared with micelles, mixed micelles with Cur~ (3-) electrostatic interaction but weak hydrophobic interaction is stronger, the Cur~ (3-) are more likely to enter the mixed micellar palisade layer. Therefore, the mixed micelles can effectively isolate Cur~ (3-) and isolated from the external environment and inhibit its degradation (2) study. With twelve alcohol and glucose Gemini surfactant (Gemini-C12) lipid vesicles as main wall material of bubble of catechin ester carrying capacity and antioxidant activity were improved. Results show that catechin ester global vertical positioning way take double molecular layer in the capsule, so it's in vesicles in effective load high load. In a certain range of catechin ester can improve the vesicle bilayer ordering and phase transition temperature. When more than this range, catechin ester loading increased the liquidity of the bilayer membrane and decrease the thermal stability. The vesicle encapsulation. Lipid peroxidation ability of catechin esters in liposome in the model is higher than that of free catechin, which may be related to catechin ester tended to locate in the phospholipid bilayer. (3) modified by electrostatic method construction of sodium carboxymethyl cellulose (CMC-Na) coated vesicles, and study the role of CMC-Na coated capsule to improve the global stability. The results showed that CMC-Na and vesicle binding was a spontaneous endothermic process driven by entropy. By controlling the concentration of CMC-Na can be obtained in small size, stable properties of coated vesicles, predicting the optimum concentration of CMC-Na and the empirical model and obtained the CMC-Na saturation concentration is consistent with the modified.CMC-Na increased the stability of pH vesicles, thermal stability and salt stability, and improve the retention rate of the core material of catechin ester under heat treatment conditions. (4) of Gemini-C12 and heat denatured ovalbumin (OVA) and to explore the interaction For the Gemini-C12 concentration of Gemini-C12-OVA composite coating of wheat gliadin (gliadin) mechanism characteristics of nanoparticles. The results show that when the Gemini-C12 OVA-Gemini-C12 mixed solution concentration is lower than the critical micelle concentration (CMC), free Gemini-C12 tends to be adsorbed on particle water interface. At this point, Gemini-C12 can increase the nucleation rate of anti solvent process in, so as to reduce the size of gliadin nanoparticles; on the other hand, it can fill the void left by OVA particle surface adsorption layer, thereby improving the storage stability of nano particles and re dispersion. When the concentration of Gemini-C12 is higher than CMC, in the positioning in the micelles rather than adsorption at the solid-liquid interface surface active agent the molecular tendency, thus the size of gliadin nanoparticles increased and reduced its storage stability and redispersibility. (5) combining with Maillard reaction and isoelectric point heating gel method Ovalbumin dextran nanogels were prepared and evaluated nano gel entrapment can improve the bioavailability of curcumin. Ovalbumin dextran nanogels were mainly spherical structure, the pH stability, storage stability and redispersibility were good. The load of curcumin nano gel by pH driving method. Nano gel entrapment in curcumin after physicochemical properties have no obvious change. Ovalbumin dextran nanogels and ovalbumin nanoparticles encapsulated curcumin can improve the conversion rate of digestion in the simulation (transformation) and bioavailability rate (bioaccessibility). The nano gel entrapment ratio of curcumin nanoparticles encapsulated curcumin conversion between the two and higher bioavailability was no significant difference, and this may be in simulating digestion similar physical and chemical properties. (6) were prepared by free radical grafting With ovalbumin and catechin covalent and studied ovalbumin and three covalent conjugates of fish oil emulsion stability. Due to the introduction of phenolic hydroxyl groups, free radical scavenging ability of ovalbumin was improved. Catechin grafting changed two level structure of ovalbumin, and increased its surface hydrophobicity and three conformational flexibility. Compared with the ovalbumin fish oil emulsion, lipid droplets were combined with smaller fish oil emulsion of ovalbumin - catechin covalently, better storage stability, lower viscosity and higher oxidation stability. Overall, oxidation stability under the condition of pH 3.5 oil emulsion than pH 7 under the condition of high, this may be related to with pH3.5 under the condition of positively charged lipid droplets of transition metal repulsion.

【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：TS201.21;O647.2
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本文编号：1518788