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分段盐析联合两步柱层析纯化巢湖蓝藻藻蓝蛋白
苏雨,张发宇,余金卫,汪家权,许子牧
0
(合肥工业大学资源与环境工程学院,合肥 230009;合肥工业大学资源与环境工程学院,合肥 230009; 合肥市东方美捷分子材料技术有限公司,合肥 230009)
摘要:
以巢湖打捞的新鲜蓝藻藻泥为处理对象,利用冻融破壁的方式获得藻蓝蛋白(phycocyanin,PC)粗提液。采取两步盐析初步纯化粗提液获得PC,再用纤维素(Cellufine A-500)阴离子交换树脂中度纯化PC,最后用羟基磷灰石(HA)柱高度纯化PC。结果表明,在室温25℃下,两步盐析(NH4)2SO4的最佳摩尔浓度分别为1.0 mol·L-1和1.8 mol·L-1,PC纯度(A620/A280)达到2.168,PC回收率可达37.5%。再经Cellufine A-500和HA两步柱层析纯化,PC纯度(A620/A280)分别达到3.211和4.133,回收率分别为20.3%和7.2%,纯度达到试剂级纯度要求。
关键词:  藻蓝蛋白  两步盐析  阴离子交换  羟基磷灰石
DOI:10.13610/j.cnki.1672-352x.20180620.009
基金项目:国家“十二五”重大科技专项项目(2012ZX07103-004)资助。
Purification of phycocyanin from Chaohu cyanobacteria by fractional salting out and two steps column chromatography
SU Yu,ZHANG Fayu,YU Jinwei,WANG Jiaquan,XU Zimu
(School of Resources and Environmental Engineering, Hefei University of Technology, Hefei 230009;School of Resources and Environmental Engineering, Hefei University of Technology, Hefei 230009; Hefei Dongfangmeijie Molecular Materials Technology Co.Ltd., Hefei 230009)
Abstract:
The fresh algae mud from Chaohu was used to obtain the crude extract of phycocyanin (PC) using freezing and thawing. The crude extract was purified by two-step salting-out, and the PC was purified by cellulose (Cellufine A-500) anion exchange resin and finally purified by hydroxyapatite (HA) column. The results showed that At room temperature (25℃), the optimum molar concentrations of two-step salting-out (NH4)2SO4 were 1.0 mol·L-1 and 1.8 mol·L-1, respectively. The PC purity (A620 / A280) reached to 2.168 and the PC recovery was 37.5 %. The purity of PC (A620 / A280) reached 3.211 and 4.133, respectively, and the recovery was 20.3% and 7.2%, respectively, which reaches the reagent grade purity.
Key words:  phycocyanin  two-step salt precipitation  anion exchange  hydroxyapatite

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