【作者】 王志忠；

【导师】 李博生；

【作者基本信息】 内蒙古农业大学 ， 植物学， 2005， 硕士

【摘要】 本文首先对螺旋藻多糖的提取工艺进行了优化,用正交试验方法对提提取温度、取液浓度、提取时间设置三水平进行了筛选研究。并用单因素试验的方法对螺旋藻多糖提取过程中的 TCA 用量,各环节的离心速度进行了研究。结果表明选用在室温下用 0.1mol/L NaOH 提取 1 小时这一处理组合的显著性最好,用这一组合去提取多糖可使提取率达到 5.195%,并且,粗多糖中多糖含量达到 99%,这一组合不仅节省了很多能量,而且使提取率超过其他提取工艺。在 TCA 用量试验中,筛选出了 TCA 的最佳用量,即每克藻粉的提取液中应加入 0.6 mL 25%TCA,从以往的 TCA 定性加入提高到定量使用,避免了蛋白质沉淀不完全或 TCA 加入过量问题的出现,确保了多糖的质量并且控制了提取成本。在离心速度试验中,分别对三次离心操作(粗提液中的非蛋白多糖物质沉淀、蛋白沉淀、多糖沉淀)的离心速度进行了筛选,最终分别选出了 4000rpm、6000 rpm、6000 rpm 三个速度。 然后对内蒙古钝顶螺旋藻(S1)与从非洲乍得湖引进的钝顶螺旋藻(S2)和从墨西哥 TEXCOCO 湖引进的极大螺旋藻(S3)以及从内蒙古钝顶螺旋藻(S1)经辐射后选育出来的抗辐射藻株(S1K)这四个藻种的所含多糖的提取分离、纯化进行了比较研究。结果表明,四个藻种的多糖含量顺序分别为 S1K 〉S1 〉S2 〉S3,而四个螺旋藻样本的完整细胞的辐射抗性的大小顺序为 S1K 〉S1 〉S2 〉S3,所以由此表明螺旋藻的多糖含量与其辐射抗性呈显著的正相关,即多糖含量越高,抗辐射能力越强。另外用凝胶层析法对四种螺旋藻所含多糖进行了分离,S1K 和 S1 均由 5 种相同的多糖组成,只是 S1k有三种多糖的含量大于 S1,这也证明了 S1K的辐射抗性大于 S1的事实;S2 含有三种多糖,S3 含有四种多糖。并对洗脱峰面积和每一峰的多糖浓度的相关性作了分析,证明洗脱峰面积和每一峰的多糖浓度之间存在着正相关关系。更多还原

【Abstract】 We have optimized the process to separate polysaccharide from Spirulina with orthogonal experiment in which we set three factors and each factor has three levels and selected the optimal amount of TCA added and centrifugal velocity In the different courses of the separation with the single-factor experiment. The results shows that the treatment adding 0.1M NaOH into the powder of Spirulina in term of the ratio of 1:7 of quality with volume while reacting for one hour can make the ratio of separation of polysaccharide reach 5.195% and the maximal purity of polysaccharide reach 99%. This group of treatment can not only save a great large of energy ,but also make the ratio of separation of polysaccharide higher than other processes to separate polysaccharide from Spirulina. We have selected optimal amount of TCA added that is 0.6 mL 25% TCA in the liquid of separation of per gram dry powder of Spirulina, so that we make the method adding TCA change from the qualitative method to the quantificational method and avoid the problem that the protain cannot be sedimentated incompletely and that we add TCA excessively , ulteriorly we insure the polysaccharide quality and reduce the cost.In addition,we also have selected the optimal centrifugal velocity in several important centrifugal stages that are 4000rpm,6000 rpm and 6000 rpm. Comparison studies on separating and sublimating polysaccharide from Spirulina platensis from Inner Mongolia(S1) and the imported S. platensis from Africa(S2) and S. maxima from Mexico(S3) and another species(S1K) that is chosen and cultivated from S1 that has been treated with 60Coγradial. The results show:the order of separation rate of polysaccharide from the four Spirulina species is S1k 〉S1 〉S2 〉S3,while the order of resistibility to radialization for them is S1k 〉S1 〉S2 〉S3, so the separation rate of polysaccharide is correlated to resistibility to radialization;S1K and S1 consist of the same five kind of polysaccharide in which the contents of three kinds of polysaccharide of S1k are larger than those of S1, while S2 and S3 consist of three and four kinds of polysaccharide respectively. Additionally,we also analyze and discuss the pertinence of the area and concentration of each composition.The result shows they are correlative positively.更多还原