【作者】 刘艳辉；

【导师】 汪志平；

【作者基本信息】 浙江大学 ， 生物物理学， 2005， 硕士

【摘要】 螺旋藻是丝状的多细胞蓝藻,是当前国内外研究与开发规模最大的经济微藻。藻胆蛋白是螺旋藻中含有藻胆素、具有荧光特性的一类生理活性蛋白,具有很高的理论研究与开发利用价值。藻胆蛋白不仅可作为天然色素用于食品、化妆品和染料等行业,而且因其具有提高机体免疫力、抗肿瘤、抗辐射、抗衰老和防治贫血等功效而广泛用于医药保健领域。同时,利用其特有的光化学活性制成的荧光分子探针和光敏剂,在临床医学诊断、免疫化学、生物工程及肿瘤激光治疗等领域也有很好的应用前景。 为筛选到能应用于生产实际的高产藻蓝蛋白的钝顶螺旋藻优良新品系,我们对Sp-S、Sp-T、Sp-YE15、Sp-CH32、Sp-HUF01、Sp-IDF和Sp-YC09这7株在分类学上有代表性、且在养殖生产中表现优良的钝顶螺旋藻品系的光合色素作了较系统的比较研究。结果发现,Sp-CH32的叶绿素和类胡萝卜素与其它螺旋藻株的相当,而藻胆蛋白含量高达细胞干重的29.994%,比上述其它6株中最高者Sp-S的高20.9%,比上述7株螺旋藻藻胆蛋白含量的平均值高36.9%。进一步测得取自工厂化养殖中6个生产批次Sp-CH32的藻胆蛋白含量在29.872～35.821%之间,平均含量为32.412%。从而表明,Sp-CH32在实验室和大规模养殖中均具有稳定、高效表达藻胆蛋白的优良特性。 针对Sp-CH32藻胆蛋白的含量高、细胞较其它藻株易于破碎等特点,我们在总结前人工作的基础上,通过研究建立了Sp-CH32藻胆蛋白的制备工艺,并对纯化出的藻蓝蛋白和别藻蓝蛋白的光谱特性等作了研究。藻体细胞经冻融破碎,所得粗提液经（NH₄）₂SO₄沉淀所得45%～50%饱和度的级份,直接上Sephacryl S-200凝胶过滤层析柱,不仅使藻胆蛋白的纯度值提高到3左右,还去除了（NH₄）₂SO₄,省去繁琐、更多还原

【Abstract】 Spirulina platensis is multi-cell filamentous blue-green alga, simultaneously, it is an economic microalga with the largest scale of research and exploiture inland and abroad. The phycobiliproteins are kinds of active protein which extensively exists in spirulina, and it bears phycobilin with fluorescence idiosyncrasy, so the phycobiliproteins possesses great values in theory research as well as exploiture. The phycobiliproteins can not only be used as natural pigments in food, cosmetic and dye industries, but also be applied to the area of medicaments and healthy foods since it can improve immune capability and resist tumour, radiation, caducity, anaemia and so on. In addition, as fluorescence molecular probe and photosensitive reagent, it has fine prospect in clinical diagnoses, immunochemistry, bioengineering and tumor laser therapy.In order to screen out the excellent Spirulina platensis strains with high-yield phycocyanin in large-scale cultivation, The contents of photosynthetic pigments of seven strains, named Sp-S, Sp-T, Sp-YE15, Sp-CH32, Sp-HUF01, Sp-IDF and Sp-YC09, which have representation in taxology and exhibit eximious quality in large-scale production, were compared. The results indicated that the contents of chlorophyll and carotenoid in Sp-CH32 were as much as the other strains’, whereas the content of phycobiliproteins achieve 29.994% of dry weight of cells, the content of phycobiliproteins in Sp-CH32 was 20.9% higher than that in Sp-S which is highest in the other six strains and 36.9% higher than the average value of the content of phycobiliproteins in seven strains. Furthermore, the contents of phycobiliproteins of six different batchs Sp-CH32 in large-scale cultivation, were between 29.872% and 35.821%, and their average content is 32.412%. The results indicated that Sp-CH32 has excellent characteristic of stable and high expression phycobiliproteins not only in laboratory but also in large-scale cultivation.On the basis of the former works and the characteristics of high-content phycobiliproteins and cells fragility in Sp-CH32, we established a preparation technology更多还原