藜麦多糖分离纯化、结构鉴定及免疫活性研究
发布时间：2021-07-21 08:03
藜麦原产于南美洲安第斯山区,具有很高的食用价值,是目前公认的适宜人类食用的“全营养食品”。近些年才开始在全球其他地区得到推广和种植,我国目前已经在山西、西藏、四川、青海等地实现规划化种植。静乐自2011年试种成功以来发展迅速,被称为“中国藜麦之乡”。本文以山西静乐藜麦为原料,对其中的多糖进行分离纯化,采用多种方法分析了多糖结构与分子形貌,研究其抗氧化活性与体内免疫活性,从而为开发利用藜麦多糖提供了理论依据。（1）采用酶解协同超声波联合方法提取藜麦中多糖,以多糖提取率为指标,经试验确定最佳辅助酶为纤维素酶,最优添加量为3%。在单因素试验的基础上,进行响应面试验,结果表明,藜麦多糖最优提取工艺为:超声温度65℃、超声时间18 min、料液比1:33（g/mL）,此时藜麦多糖的提取率为68.08%,与理论值70.78%接近。酶具有量少而催化效率高的特点,能够加速多糖溶出,有效降低生产成本;超声波产生空化作用,使细胞破碎,细胞内溶物更好地溶出。酶法辅助超声波提取法可以明显提高藜麦多糖的提取率,通过对比试验表明,经酶法辅助超声波提取比单一采用超声波提取,多糖提取率增加1.5倍。（2）上述最优条件...
【文章来源】：山西大学山西省
【文章页数】：79 页
【学位级别】：硕士
【文章目录】：
中文摘要
Abstract
Chapter 1 Review of the literature
1.1 Introduction of Chenopodium quinoa Willd
1.1.1 Botanical description
1.1.2 Analysis of Nutritional Value
1.1.3 physiology function
1.2 Introduction of polysaccharides
1.2.1 Extraction, separation and purification of polysaccharides
1.2.2 Structure and analysis of polysaccharides
1.2.3 Conformational features of polysaccharide
1.3 Research on the biological activity of polysaccharides
1.3.1 Immunomodulatory activities
1.3.2 Oxidation resistance
1.3.3 Antitumor activities
1.3.4 Hypoglycemic activity
1.4 Research Meaning
Chapter 2 Synergistic Extraction of Chenopodium quinoa Polysaccharide by Enzyme-Ultrasound
2.1 Introduction
2.2 Materials and methods
2.2.1 Materials and reagents
2.2.2 Instruments and equipment
2.2.3 Measurement of polysaccharide concentration and extraction yield
2.2.4 Extraction of polysaccharides from quinoa
2.2.5 Determination of the optimal amount of coenzyme and its addition
2.2.6 Optimization of ultrasonic extraction conditions
2.2.7 Statistical analysis
2.3 Results and analysis
2.3.1 The best auxiliary enzyme and its dosage
2.3.2 Results of single-factor tests
2.3.3 Results of central composite design
2.3.4 Validation experiments
2.4 Conclusions
Chapter 3 The separation and Purification of CPS
3.1 Introduction
3.2 Materials and methods
3.2.1 Materials and reagents
3.2.2 Removal of starch
3.2.3 Detection of protein concentrations
3.2.4 Deproteinization by the enzymatic combined with Sevag method
3.2.5 DEAE-52 cellulose chromatography
3.2.6 Sephadex G-50 gel chromatography
3.3 Results and analysis
3.3.1 Deproteinization of quiona polysaccharides
3.3.2 Classification of polysaccharides
3.4 Conclusions
Chapter 4 Structural elucidation of QPS1
4.1 Introduction
4.2 Materials and methods
4.2.1 Materials and instruments
4.2.2 Determination of molecular weight
4.2.3 HPLC analysis for monosaccharide composition
4.2.4 IR spectra and 1HNMR analysis
4.2.5 Congo red assay
4.2.6 Observation of molecular and apparent morphology
4.3 Results and analysis
4.3.1 Molecular weight and homogeneity detection
4.3.2 Analysis of monosaccharide composition
4.3.3 IR analysis of QPS1
4.3.4 NMR analysis of QPS1
4.3.5 Analysis of the triple helix structure
4.3.6 Atomic force microscopy (AFM)
4.3.7 Scanning electron microscopy
4.4 Conclusions
Chapter 5 Study on antioxidant activity in vitro and Immunomodulatory activitiesin vivo
5.1 Introduction
5.2 Materials and methods
5.2.1 Materials and reagents
5.2.2 Study on antioxidant activity in vitro
5.2.3 Inhibition of α-amylase activity
5.2.4 Animal experiment design
5.2.5 Organ index assay
5.2.6 Cytokine secretion and Serum lysozyme activity
5.2.7 Monocyte-macrophage phagocytosis assay
5.2.8 Delayed type hypersensitivity assay
5.2.9 Statistical analysis
5.3 Results and analysis
5.3.1 Trolox standard curve
5.3.2 ABTS and DPPH radical scavenging activity
5.3.3 ·OH radical scavenging activity
5.3.4 Inhibition effects of QPS on α-amylase
5.3.5 Changes in body weight during feeding
5.3.6 Effect of QPS1 on immune organs
5.3.7 Effect of QPS1 on serum levels of cytokines and LYSO inimmunosuppressed mice
5.3.8 Effect of QPS1 treatment regulated mononuclear macrophage function
5.3.9 Effect of QPS1 treatment on delayed hypersensitivity
5.4 Conclusions
Chapter 6 Conclusion and prospect
6.1 Conclusion
6.2 Prospects
References
攻读学位期间取得的研究成果
致谢
个人简历及联系方式
【参考文献】：
期刊论文
[1]羊栖菜多糖提取条件优化及其抗氧化活性的研究[J]. 刘洪超,应苗苗,周雨暪,杨靖亚,汪之和,施文正. 食品工业科技. 2017(06)
本文编号：3294642
【文章来源】：山西大学山西省
【文章页数】：79 页
【学位级别】：硕士
【文章目录】：
中文摘要
Abstract
Chapter 1 Review of the literature
1.1 Introduction of Chenopodium quinoa Willd
1.1.1 Botanical description
1.1.2 Analysis of Nutritional Value
1.1.3 physiology function
1.2 Introduction of polysaccharides
1.2.1 Extraction, separation and purification of polysaccharides
1.2.2 Structure and analysis of polysaccharides
1.2.3 Conformational features of polysaccharide
1.3 Research on the biological activity of polysaccharides
1.3.1 Immunomodulatory activities
1.3.2 Oxidation resistance
1.3.3 Antitumor activities
1.3.4 Hypoglycemic activity
1.4 Research Meaning
Chapter 2 Synergistic Extraction of Chenopodium quinoa Polysaccharide by Enzyme-Ultrasound
2.1 Introduction
2.2 Materials and methods
2.2.1 Materials and reagents
2.2.2 Instruments and equipment
2.2.3 Measurement of polysaccharide concentration and extraction yield
2.2.4 Extraction of polysaccharides from quinoa
2.2.5 Determination of the optimal amount of coenzyme and its addition
2.2.6 Optimization of ultrasonic extraction conditions
2.2.7 Statistical analysis
2.3 Results and analysis
2.3.1 The best auxiliary enzyme and its dosage
2.3.2 Results of single-factor tests
2.3.3 Results of central composite design
2.3.4 Validation experiments
2.4 Conclusions
Chapter 3 The separation and Purification of CPS
3.1 Introduction
3.2 Materials and methods
3.2.1 Materials and reagents
3.2.2 Removal of starch
3.2.3 Detection of protein concentrations
3.2.4 Deproteinization by the enzymatic combined with Sevag method
3.2.5 DEAE-52 cellulose chromatography
3.2.6 Sephadex G-50 gel chromatography
3.3 Results and analysis
3.3.1 Deproteinization of quiona polysaccharides
3.3.2 Classification of polysaccharides
3.4 Conclusions
Chapter 4 Structural elucidation of QPS1
4.1 Introduction
4.2 Materials and methods
4.2.1 Materials and instruments
4.2.2 Determination of molecular weight
4.2.3 HPLC analysis for monosaccharide composition
4.2.4 IR spectra and 1HNMR analysis
4.2.5 Congo red assay
4.2.6 Observation of molecular and apparent morphology
4.3 Results and analysis
4.3.1 Molecular weight and homogeneity detection
4.3.2 Analysis of monosaccharide composition
4.3.3 IR analysis of QPS1
4.3.4 NMR analysis of QPS1
4.3.5 Analysis of the triple helix structure
4.3.6 Atomic force microscopy (AFM)
4.3.7 Scanning electron microscopy
4.4 Conclusions
Chapter 5 Study on antioxidant activity in vitro and Immunomodulatory activitiesin vivo
5.1 Introduction
5.2 Materials and methods
5.2.1 Materials and reagents
5.2.2 Study on antioxidant activity in vitro
5.2.3 Inhibition of α-amylase activity
5.2.4 Animal experiment design
5.2.5 Organ index assay
5.2.6 Cytokine secretion and Serum lysozyme activity
5.2.7 Monocyte-macrophage phagocytosis assay
5.2.8 Delayed type hypersensitivity assay
5.2.9 Statistical analysis
5.3 Results and analysis
5.3.1 Trolox standard curve
5.3.2 ABTS and DPPH radical scavenging activity
5.3.3 ·OH radical scavenging activity
5.3.4 Inhibition effects of QPS on α-amylase
5.3.5 Changes in body weight during feeding
5.3.6 Effect of QPS1 on immune organs
5.3.7 Effect of QPS1 on serum levels of cytokines and LYSO inimmunosuppressed mice
5.3.8 Effect of QPS1 treatment regulated mononuclear macrophage function
5.3.9 Effect of QPS1 treatment on delayed hypersensitivity
5.4 Conclusions
Chapter 6 Conclusion and prospect
6.1 Conclusion
6.2 Prospects
References
攻读学位期间取得的研究成果
致谢
个人简历及联系方式
【参考文献】：
期刊论文
[1]羊栖菜多糖提取条件优化及其抗氧化活性的研究[J]. 刘洪超,应苗苗,周雨暪,杨靖亚,汪之和,施文正. 食品工业科技. 2017(06)
本文编号：3294642
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