【作者】 王芙蓉；

【导师】 梁运祥；

【作者基本信息】 华中农业大学 ， 微生物学， 2007， 硕士

【摘要】 花粉是植物雄性配子体，存在于雄性花粉囊内，不仅具有低脂肪、高蛋白的特点，而且含有人体生存所需的多种营养物质，素有“微型营养库”之称，在国际上被誉为“完全营养品”，尤其是其中多糖、黄酮两类化合物，在增强机体免疫力、降血脂、抗肿瘤、抗氧化等方面都有显著的功效。本实验以油菜花粉为原料，研究了其破壁方法及多糖、黄酮两种活性物质的提取与纯化工艺。研究结果表明：(1)采用酶解与超声波相结合的方法对油菜花粉进行破壁处理，具体步骤为：1g花粉与10ml混合酶液配成10％的花粉溶液，其中果胶酶酶活约600U，纤维素酶酶活约450U，于45℃，pH4.5的条件下酶解12h，再用超声波破碎仪在400W条件下处理10min，破壁率最终可达约90％以上。(2)采用水提法对油菜花粉多糖进行提取的最佳条件为：在90℃水浴条件下，按1∶8的固液比提取2h，并重复操作一次。按最佳条件分别对破壁与未破壁花粉进行提取，多糖得率分别为1.631％和1.256％，前者是后者的1.3倍。采用TCA法去蛋白和10倍乙醇沉淀后，粗多糖中多糖含量约20％。再经乙醇分级，得到三个级分，分别为PPA、PPB、PPC，SephadexG-200柱层析显示PPA为非均一多糖，PPB和PPC为均一多糖。(3)采用乙醇提取法对油菜花粉黄酮提取的最佳条件为：在95℃水浴条件下，按1∶35的固液比提取2h。在最优条件下，对破壁与未破壁花粉进行提取，黄酮得率分别为3.167％和2.103％，前者是后者的1.5倍。干燥后粗提物中黄酮的含量为18.6％，经大孔吸附树脂DM-130纯化后黄酮含量可提高到60.5％。更多还原

【Abstract】 Pollen is the male gametophyte of gymnosperm and angiosperm. It’s not onlylow fat and high protein, but also contains many kinds of nutrient which are necessaryto human body. It is well known as a natural nutrition, health food and "Perfect food".The anti-tumor activity of pollen had attracted more attention in food, biochemicalland medical fields. Using Rape pollen as the raw material in this research, themethod for breaking wall and for extracting and purifying polysaccharides andflavonoids were investigated. The results show:(1)The wall was broken by enzymolysis and supersonic, lg pollen withl0mlmixed enzyme solutions, in which pectolytic activity 600U and cellulose activity450U. The optimum conditions were: enzymolysis at 45℃and pH4.5 for 2 hours andthen supersonic at 400W for 10min. The wall-broken efficiency can reach above 90％.(2)The polysaccharides of Rape pollen were extracted with water. The optimumconditions of extraction were 1:8 solid-liquid ratio at 90℃for 2 hours by 2times. Theextraction percent of polysaccharides from wall-broken pollen was 1.631％, whichwas 30％more than unbroken pollen. After deproteinization and sedimentation, thecontent of polysaccharides can reach 20％. We got there fractions after the alcohol:PPA、PPB and PPC. It shows that the PPB and PPC are homogeneous polysaccharidesthrough Sephadex G-200, but PPA is not homogeneous polysaccharides.(3) The flavonoids of Rape pollen were extracted with ethanol, and the extractionwas purified with macroporous adsorption resin. The optimum conditions ofextraction were 95％ethanol and 1:35 solid-liquid ratio at 95℃for 2 hours. Theextraction percent of flavonoids from wall-broken pollen was 3.167％, which was50.6％more than unbroken pollen. DM-130 was selected as the optimum resin of 8types of macroporous adsorption resin. When the resin was loaded with 16.78mgextraction and desorbed with 50ml 50％ethanol, the content of flavonoids could reach60.5％.更多还原