钝顶螺旋藻藻蓝蛋白的提取工艺优化及其纯化研究  

Study on the Optimization of the Extraction and Purification of Phycocyanin from Spirulina platensis

作　　者：陈盛仕 刘梓宜 李梦琪 米顺利 陈显强 张玲[1,3] CHEN Shengshi;LIU Ziyi;LI Mengqi;MI Shunli;CHEN Xianqiang;ZHANG Ling(Guangxi Scientific Research Center of Traditonal Chinese Medicine,Nanning,Guangxi,530200,China;Institute of Marine Drugs,Guangxi University of Chinese Medicine,Nanning,Guangxi,530200,China;Guangxi Key Laboratory of Chinese Medicine Foundation Research,Guangxi University of Chinese Medicine,Nanning,Guangxi,530200,China)

机构地区：[1]广西中医药大学科学试验中心,广西南宁530200 [2]广西中医药大学海洋药物研究院,广西南宁530200 [3]广西中医药大学,广西中医基础研究重点试验室,广西南宁530200

出　　处：《广西科学院学报》2022年第4期388-402,共15页Journal of Guangxi Academy of Sciences

基　　金：广西科技计划项目重点研发计划(桂科AB19110014);广西中医药大学一流学科建设开放课题(2019XK110);广西中医药大学海洋药物研究院团队科研专项经费项目(2018ZD005-C04);广西中医药大学中医基础研究重点实验室自主研究课题(K201137903,16-380-58-05)资助。

摘　　要：以钝顶螺旋藻(Spirulina platensis)藻蓝蛋白(Phycocyanin)的纯度和回收率为指标,考察溶胀法和冻融法中不同破壁影响因素及活性炭吸附法中活性炭目数、活性炭加入量和吸附时间对藻蓝蛋白粗提的影响,并结合响应面分析法优化提取工艺参数,得到最优破壁条件和活性炭吸附条件。结果表明,溶胀法最优提取条件为料液比1∶50(g/mL),溶胀时间24 h,溶胀次数2次,测得藻蓝蛋白纯度为0.47,得率为2.17%;冻融法最优提取条件为料液比1∶50(g/mL),冷冻时间8 h,冻融次数2次,测得藻蓝蛋白纯度为0.34,得率为2.19%。活性炭吸附法最优工艺条件为300目活性炭,活性炭加入量0.7 g/20 mL,吸附时间10 min,藻蓝蛋白纯度为0.80,回收率为73.23%。疏水层析法纯化得到食品级藻蓝蛋白(纯度P≥0.7)和医药级藻蓝蛋白(纯度P≥3.0),总回收率为61.15%。因此,溶胀-活性炭吸附-疏水层析三步提纯藻蓝蛋白工艺合理可行,实现了医药级藻蓝蛋白的分离提纯,降低了工业生产成本,具有实用价值。The purity and recovery rate of phycocyanin from Spirulina platensis were used as indexes to investigate the effects of different wall-breaking factors in swelling method and freeze-thaw method,as well as the number of activated carbon mesh,activated carbon addition and adsorption time in the activated carbon adsorption method on the crude extraction of phycocyanin.The extraction process parameters were optimized by response surface methodology,and the optimal wall-breaking conditions and activated carbon adsorption conditions were obtained.The results showed that the optimum extraction conditions of swelling method were as follows:Solid-liquid ratio was 1∶50(g/mL),swelling time was 24 h,and swelling times was 2.The purity of phycocyanin was 0.47 and the yield was 2.17%.The optimum extraction conditions of freeze-thaw method were:Solid-liquid ratio 1∶50(g/mL),freezing time 8 h,freeze-thaw times 2 times.The purity of phycocyanin was 0.34 and the yield was 2.19%.The optimum conditions of activated carbon adsorption method were as follows:300 mesh activated carbon,0.7 g/20 mL activated carbon,adsorption time of 10 min.The purity of phycocyanin was 0.80 and the recovery rate was 73.23%.Food grade phycocyanin(purity P≥0.7)and pharmaceutical grade phycocyanin(purity P≥3.0)were purified by hydrophobic chromatography,and the total recovery rate was 61.15%.Therefore,swelling-activated carbon adsorption-hydrophobic chromatography three-step purification of phycocyanin process is reasonable and feasible,which can realize the separation and purification of pharmaceutical-grade phycocyanin,reduce industrial production costs and has a practical value.

关 键 词：钝顶螺旋藻 藻蓝蛋白 溶胀法 活性炭 响应面法优化 疏水层析 

分 类 号：Q816[生物学—生物工程]