盐析法联合柱层析法纯化藻蓝蛋白的研究

论文摘要

本研究采用分段盐析法和柱层析法从巢湖新鲜蓝藻中提取纯化藻蓝蛋白。首先,使用硫酸铵、柠檬酸三铵、柠檬酸钠和硫酸钠对藻蓝蛋白粗提液进行两步分段盐析纯化,并利用紫外-可见分光光度法对藻蓝蛋白溶液进行纯度及回收率分析,以此确定了硫酸铵、柠檬酸三铵、柠檬酸钠和硫酸钠的最佳用量。其结果为:硫酸铵、柠檬酸三铵、柠檬酸钠和硫酸钠在第一和第二盐析过程中的最佳摩尔浓度分别为:硫酸铵（NH4）2SO4为1.0 mol/L和1.7 mol/L;柠檬酸三铵C6H17N3O7为0.7mol/L和1.3 mol/L;柠檬酸钠C6H5Na3O7为0.5 mol/L和0.9 mol/L;柠檬酸钠C6H17N3O7为1.0 mol/L和1.3 mol/表示硫酸钠Na2SO4。经过两步分段盐析,发现随着无机盐摩尔浓度的增高,杂质去除量增大到一定程度后会去除藻蓝蛋白,故纯度和收率都先提高再降低。最优结果表明,使用（NH4）2SO4纯化后,藻蓝蛋白纯度达2.0以上,藻蓝蛋白回收率较高。为了进一步提高藻蓝蛋白的纯度,用（NH4）2SO4进行第二次盐析后,利用离子交换色谱法进一步纯化藻蓝蛋白,研究了pH、离子强度、洗脱速度与进样浓度对纯化效果的影响。其最佳条件为:纤维素A-500,洗脱液pH值7.0,样品浓度1-2 mol/L,加样速度2cm/min,洗脱液NaCl浓度0.25mol/L,最终得到纯度在4.0以上的试剂级藻蓝蛋白。

论文目录

• Acknowledgement
• ABSTRACT
• 摘要
• Chapter 1 Introduction
•   1.1 Background information
•     1.1.1 Location of Chaohu Lake
•     1.1.2 Overview of Chaohu Lake Basin
•     1.1.3 Analysis of eutrophication status of Chaohu Lake
•     1.1.4 Current status of cyanobacteria resource utilization in Chaohu Lake
•   1.2 Research status of phycobiliprotein purification at home and abroad
•     1.2.1 Concepts related to purification of Phycobiliprotein
•     1.2.2 Research status of phycobiliprotein purification
• Chapter 2 Materials and Methods
•   2.1 Materials
•     2.1.1 Organism and culture conditions
•     2.1.2 Laboratory equipment
•     2.1.3 Experiment reagents and materials
•   2.2 Methods
•     2.2.1 Phycobiliprotein extraction and purification
•   2.3 Main analysis methods
• Chapter 3 Results and analysis
•   3.1 Ammonium sulfate precipitation
•     3.1.1 One step salting out
•     3.1.2 Two-step salting out
•   3.2 Triammonium citrate precipitation
•     3.2.1 One-step salting out
•     3.2.2 Two-step of salting out
•   3.3 Sodium citrate precipitation
•     3.3.1 One-step of salting out
•     3.3.2 Two-step of salting out
•   3.4 Sodium sulfate precipitation
•     3.4.1 One-step of salting out
•     3.4.2 Two-step of salting out
•   3.5 Analysis
• Chapter 4 Purification of Phycocyanin by ion Exchange Chromatography
•   4.1 Literature review
•   4.2 Materials and methods
•     4.2.1 Experimental Materials and Instruments
•     4.2.2 IEC precipitation methods
•   4.3 Selection of Fillers for Ion Exchange Chromatography
•   4.4 Results and analysis
•     4.4.1 Effect of eluent p H on Purification of phycocyanin by CellufineA-500
•     4.4.2 Effect of ionic strength on purification of phycocyanin by columnchromatography
•     4.4.3 Effect of elution rate on the purification of phycocyanin by columnchromatography
•     4.4.4 Effect of Sample Concentration on Purification of Phycocyanin byColumn Chromatography
•   4.5 Response Surface Experiment of Cellufine A-500 ColumnChromatography
• Chapter 5 Conclusion And Prospect
•   5.1 Conclusion
•   5.2 Prospect
• References
• List of Academic Activities and Achievements during the Degree

文章来源

类型: 硕士论文

作者: Kolawole Bienvenu Hanania E BONI

导师: 汪家权

关键词: 水华蓝藻,藻蓝蛋白,纯化,试剂级,柱层析

来源: 合肥工业大学

年度: 2019

分类: 基础科学,工程科技Ⅰ辑

专业: 生物学,环境科学与资源利用

单位: 合肥工业大学

分类号: Q51;X524

DOI: 10.27101/d.cnki.ghfgu.2019.000198

总页数: 58

文件大小: 3759K

下载量: 41