改性介孔材料分离组氨酸二肽技术研究
发布时间：2019-03-21 12:48
【摘要】：肌肽(Car,β-alanyl-L-histidine)和鹅肌肽(Ans,β-alanyl-L-1-methylhistidine)等组氨酸二肽广泛存在于多脊椎动物的骨骼肌和代谢旺盛的脑中,具有多种生理活性和功能,可作为食品添加剂应用于食品加工行业中,低值肉类尤其是下蛋母鸡等是其廉价来源。介孔氧化硅材料具有高比表面积和大小可调的孔径率在吸附和分离中具有巨大的应用潜力。本论文旨在利用组氨酸二肽含有咪唑基的特性,设计合成金属掺杂介孔材料,对其进行表征并研究其对组氨酸二肽的吸附特性,建立动力学和吸附等温线模型研究机理。结果为金属掺介孔材料在组氨酸二肽等功能肽分离纯化领域的应用提供了理论依据。首先,建立组氨酸二肽的HPLC检测方法。采用OPA柱前衍生法快速测定提取液中肌肽和鹅肌肽含量。测定不同物种及同一物种不同部位中组氨酸二肽含量,发现蛋鸡肉中含有较高的组氨酸二肽(21.58 pmol/g)。又由于蛋鸡经济价值低廉,因此是提取组氨酸二肽的理想来源。采用切丁、绞碎、搅拌、超声、蒸煮及酶解方法提取组氨酸二肽,绞碎后搅拌方法能够最大化的提取二肽且引入其他氨基酸及多肽较少。采用1000 Da超滤膜和500 Da纳滤膜两步法进行分离,除大分子蛋白和肽,经氨基酸分析,组氨酸二肽含量约60%。其次,采用两步法合成金属掺杂介孔材料。采用羧基化改性剂CES在SBA-15骨架中加入羧基,通过XRD、TEM、BET等进行表征,羧基化介孔材料S-COOH仍保持SBA-15中二维六方孔道结构,而随CES添加量增加,孔道发生破坏、塌陷和粘连,有序性降低。经改性后S-COOH具有Ⅳ型吸附曲线和H1型滞后环,而改性后孔容、孔径和比表面积均有下降。采用金属硝酸盐合成金属掺杂介孔材料,以Ni/SBA-15为例,采用XRD、 BET、ICP等进行表征发现,添加金属的浓度需在一定范围内。第三,对合成材料吸附组氨酸二肽特性进行研究。探究改性介孔材料预处理条件、金属种类、金属添加量对组氨酸二肽吸附的影响,结果表明Ni50/SBA-15具有良好的吸附容量；研究pH和无机盐对其吸附的影响,发现pH 6-7范围内具有较大的吸附容量(20℃C,当pH 7.0,添加量0.5 g/L时,吸附容量为0.69mmol/g),无机盐对其吸附无较大影响。对Ni50/SBA-15吸附组氨酸二肽洗脱的研究中发现,采用50mmol/L咪唑为最佳洗脱条件。最后,建立Ni50/SBA-15对组氨酸二肽吸附的动力学方程和吸附等温线。分别采用伪一级、伪二级和颗粒内扩散模型进行动力学拟合,发现其符合伪二级动力学方程(R2在0.983-0.999范围内)。采用Langmuir模型和F reundlich模型对其进行拟合发现,吸附过程符合Langmuir模型(R2在0.994-0.998范围内),吸附为单层吸附。由Freundlich模型得出,该吸附为可逆吸附。
[Abstract]:Histidine dipeptides, such as Car, 尾-alanyl-L-histidine and Ans, 尾-alanyl-L-1-methylhistidine, exist widely in skeletal muscle and metabolic brain of polyvertebrates, and have many physiological activities and functions. It can be used as a food additive in the food processing industry, low-value meat, especially laying hens are its cheap source. Mesoporous silica materials have great potential in adsorption and separation with high specific surface area and adjustable pore diameter. The purpose of this paper is to design and synthesize metal-doped mesoporous materials based on the properties of histidine dipeptide containing imidazolyl groups and to study the adsorption characteristics of histidine dipeptides and to establish a kinetic and adsorption isotherm model for studying the mechanism of the metal-doped mesoporous materials. The results provide a theoretical basis for the application of metal-doped mesoporous materials in the separation and purification of functional peptides such as histidine dipeptides. Firstly, a HPLC method for the detection of histidine dipeptide was established. The contents of carnosine and goose carnosine in the extract were determined by OPA pre-column derivatization. The content of histidine dipeptide in different species and different parts of the same species was determined. It was found that there was a higher histidine dipeptide (21.58 pmol/g) in egg chicken. Because the economic value of laying hens is low, it is an ideal source for extracting histidine dipeptide. Histidine dipeptides were extracted by cutting, chopping, stirring, ultrasonic, cooking and enzymolysis methods. The dipeptides were extracted maximally by agitation method and the addition of other amino acids and polypeptides was less than that of other amino acids and peptides. 1000 Da ultrafiltration membrane and 500 Da nanofiltration membrane were used for separation. The content of histidine dipeptide was about 60% after amino acid analysis. Secondly, metal-doped mesoporous materials were synthesized by two-step method. Carboxylation modifier CES was used to add carboxyl groups to SBA-15 framework and characterized by XRD,TEM,BET et al. The carboxylation mesoporous material S-COOH still maintained the two-dimensional hexagonal pore structure in SBA-15. However, with the increase of CES content, the pore channel was destroyed. Collapse and adhesion, order decreased. The modified S-COOH exhibited type 鈪，
本文编号：2444945
[Abstract]:Histidine dipeptides, such as Car, 尾-alanyl-L-histidine and Ans, 尾-alanyl-L-1-methylhistidine, exist widely in skeletal muscle and metabolic brain of polyvertebrates, and have many physiological activities and functions. It can be used as a food additive in the food processing industry, low-value meat, especially laying hens are its cheap source. Mesoporous silica materials have great potential in adsorption and separation with high specific surface area and adjustable pore diameter. The purpose of this paper is to design and synthesize metal-doped mesoporous materials based on the properties of histidine dipeptide containing imidazolyl groups and to study the adsorption characteristics of histidine dipeptides and to establish a kinetic and adsorption isotherm model for studying the mechanism of the metal-doped mesoporous materials. The results provide a theoretical basis for the application of metal-doped mesoporous materials in the separation and purification of functional peptides such as histidine dipeptides. Firstly, a HPLC method for the detection of histidine dipeptide was established. The contents of carnosine and goose carnosine in the extract were determined by OPA pre-column derivatization. The content of histidine dipeptide in different species and different parts of the same species was determined. It was found that there was a higher histidine dipeptide (21.58 pmol/g) in egg chicken. Because the economic value of laying hens is low, it is an ideal source for extracting histidine dipeptide. Histidine dipeptides were extracted by cutting, chopping, stirring, ultrasonic, cooking and enzymolysis methods. The dipeptides were extracted maximally by agitation method and the addition of other amino acids and polypeptides was less than that of other amino acids and peptides. 1000 Da ultrafiltration membrane and 500 Da nanofiltration membrane were used for separation. The content of histidine dipeptide was about 60% after amino acid analysis. Secondly, metal-doped mesoporous materials were synthesized by two-step method. Carboxylation modifier CES was used to add carboxyl groups to SBA-15 framework and characterized by XRD,TEM,BET et al. The carboxylation mesoporous material S-COOH still maintained the two-dimensional hexagonal pore structure in SBA-15. However, with the increase of CES content, the pore channel was destroyed. Collapse and adhesion, order decreased. The modified S-COOH exhibited type 鈪，
本文编号：2444945
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