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pHEMA晶胶微球的制备及啤酒酵母转化液中ATP的分离

ISSN号：1003-9015
期刊名称：高校化学工程学报
时间：2012.12
页码：119-124
分类：TQ425.232[化学工程] TQ028.8
作者机构：[1]浙江工业大学化学工程与材料学院绿色合成技术国家重点实验室培育基地,浙江杭州310032, [2]中南大学资源加工与生物工程学院,湖南长沙410083, [3]中国地质大学(武汉)机械与电子信息学院,湖北武汉430074, [4]浙江大学流体动力与机电系统国家重点实验室,浙江杭州310027
相关基金：国家自然科学基金资助项目（20606031,21036005）; 科技部中欧政府间国际合作专项项目（1017）; 湖北省自然科学基金（2010CDB04101）; 流体动力与机电系统国家重点实验室开放基金项目（GZKF-201010）
相关项目：抗体分离过程中的介质、方法和集成化研究

关键词：聚甲基丙烯酸羟乙酯, 晶胶微球, 填充床, 三磷酸腺苷, 层析, poly（2-hydroxyethyl methacrylate）, cryogel microbeads, packed bed, adenosine triphosphate, chromatography

中文摘要：

利用微通道结合结晶致孔方法制备了孔隙发达、粒径分布均匀的聚甲基丙烯酸羟乙酯（pHEMA）晶胶微球。晶胶微球平均粒径924.4μm,一致性多分散系数0.21。晶胶微球填装于直径10 mm层析柱内,填充高度91 mm;经甲基丙烯酸二甲氨乙酯（DMAEMA）修饰后应用于三磷酸腺苷（ATP）分离。层析过程流速为2 cm min 1,以0.01 mol L 1HCl为缓冲液,分别用0.03 mol L 1和1 mol L 1NaCl溶液进行洗脱。利用晶胶微球填充床从三组不同ATP含量转化液中可获得纯度达93.4%~96.9%的ATP,总回收率为47.0%~82.7%。

英文摘要：

Poly（2-hydroxyethyl methacrylate） cryogel microbeads were prepared by combining the microchannel and cryo-polymerization methods under frozen condition.The mean diameter of the cryogel microbeads is 924.4 μm with a narrow particle size distribution and the consistency of multi-dispersion coefficient is 0.21.The prepared cryogel microbeads were packed in a chromatography column with diameter of 10 mm and packing height of 91 mm and then they were grafted with N,N-dimethylaminoethyl methacrylate（DMAEMA） monomer.The grafted cryogel microbead bed was used to isolate the adenosine triphosphate（ATP）.Chromatographic separation of ATP from the fermentation broths was carried out at the flow velocity of 2 cm.min.1 with 0.01 mol.L.1 HCl as running buffer and 0.03 and 1 mol.L.1 NaCl solutions,respectively as eluting liquid.The high purity ATP was recovered from the Saccharomyces cerevisiae fermentation broth feedstocks containing different contents of ATP,adenosine diphosphate（ADP）,adenosine monophosphate（AMP）,other impurities and yeast cells.The purities and the recovery ratios of the obtained ATP are about 93.4%~96.9% and 47.0%~82.7%,respectively.