条斑紫菜藻胆蛋白提纯方法优化探索  

Optimization of Method for Phycobiliprotein Extraction and Purification from Porphyra yezoensis

作　　者：蔡春尔[1] 周铭[1] 李春霞[1] 柳俊秀[1] 汪卿[1] 何培民[1] 

机构地区：[1]上海海洋大学水产与生命学院海洋生物系,上海200090

出　　处：《生物技术通报》2009年第2期98-102,共5页Biotechnology Bulletin

基　　金：国家863计划项目(2006AA10Z323;2007AA09Z406);上海市科委"登山行动计划"项目(06dz12015);上海市教委重点项目(07ZZ135);上海高校选拔培养优秀青年教师科研专项基金(科07-13);上海市重点学科建设项目(Y1101)

摘　　要：采用溶胀法与组织捣碎法相结合的紫菜叶状体细胞破碎方法,研究了不同物液比、缓冲液浸泡时间和硫酸氨盐析次数对藻胆蛋白纯度和产率的影响,对比了各种羟基磷灰石的层析效果,并对所得藻红蛋白和藻蓝蛋白做了吸收光谱、荧光发射光谱和SDS-PAGE鉴定。结果表明,在物液比为1∶5,浸泡时间为36h时,紫菜综合破碎效果最佳;经过4次硫酸氨盐析后,藻红蛋白和藻蓝蛋白的纯度最高,分别达到1.71和0.98,采用Siegelman的方法制备的羟基磷灰石一次层析所得藻红蛋白和藻蓝蛋白的纯度最高,分别达到4.73和4.42,产率分别为0.144%和0.042%,且光谱和电泳鉴定结果均达到商品化要求。The crude extract of phycobiliproteins was prepared from thallus of Porphyra yezoensis with combination of dissolving-bulging and tissue-homogenator methods. Effects of solvent volume,treating time,ammonium sulphate precipitation time,and hydroxylapatite （HA）chromatography on purity and yield of phycobiliprotein were studied. The results showed that the optimum conditions for purification,including the ratio of thallus to buffer （1：5）,treatment with dissolving-bulging,precipitation with ammonium sulphate for four times,all of which increased the purity of phycoerythrin and phycocyanin up to 1.7l and 0.98,respectively. Due to HAP chromatography according to Siegelrnan, the phycoerythrin and phycocyanin purity increased to 4.73 and 4.42,with the yield of 0.144% and 0.042%.

关 键 词：条斑紫菜 藻红蛋白 藻蓝蛋白 硫酸氨盐析 羟基磷灰石 

分 类 号：Q946[生物学—植物学]