Separation of cytidine triphosphate from Saccharomyces cerevisiae broth by anion exchange poly(2-hydroxyethyl methacrylate) composite cryogel embedded with SiO2 nanoparticles

doi:10.3969/j.issn.0438-1157.2014.10.026

CIESC Journal ›› 2014, Vol. 65 ›› Issue (10): 3938-3945.DOI: 10.3969/j.issn.0438-1157.2014.10.026

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Separation of cytidine triphosphate from Saccharomyces cerevisiae broth by anion exchange poly(2-hydroxyethyl methacrylate) composite cryogel embedded with SiO₂ nanoparticles

LIU Jie¹, SHEN Shaochuan¹, CHEN Ping¹, XU Linhong², YUN Junxian¹, YAO Kejian¹

1 State Key Laboratory Breeding Base of Green Chemistry Synthesis Technology, School of Chemical Engineering, Zhejiang University of Technology, Hangzhou 310032, Zhejiang, China;
2 School of Mechanical & Electrical Information, China University of Geosciences (Wuhan), Wuhan 430074, Hubei, China)

Received:2014-03-14 Revised:2014-07-07 Online:2014-10-05 Published:2014-10-05
Supported by:
supported by the International Science & Technology Cooperation Program between China and Europe Country’s Governments from the Ministry of Science and Technology of China (1017), the National Natural Science Foundation of China (21036005) and the Zhejiang Provincial Natural Science Foundation (LZ14B060001).

内嵌纳米粒阴离子交换聚甲基丙烯酸羟乙酯复合晶胶分离三磷酸胞苷

刘杰¹, 沈绍传¹, 陈平¹, 徐林红², 贠军贤¹, 姚克俭¹

1 浙江工业大学化学工程学院绿色化学合成技术国家重点实验室培育基地, 浙江杭州 310032;
2 中国地质大学（武汉）机械与电子信息学院, 湖北武汉 430074

通讯作者: 贠军贤
基金资助:
科技部对欧盟政府间国际科技合作项目（1017）；国家自然科学基金项目（21036005）；浙江省自然科学基金项目（LZ14B060001）。

Abstract

Abstract: An anion exchange poly(2-hydroxyethyl methacrylate) composite cryogel embedded with SiO₂ nanoparticles was prepared by cryo-polymerization followed by graft polymerization with monomer N,N-dimethylaminoethyl methacrylate. This cryogel was used to isolate cytidine triphosphate from Saccharomyces cerevisiae transformation broth. Chromatographic separation was performed with 0.01 mol·L^-1 HCl as running buffer together with 0.02 mol·L^-1 and 0.1 mol·L^-1 NaCl solutions (prepared in running buffer) as two-step elution liquids. Cytidine triphosphate purity was analyzed by high performance liquid chromatography. Cytidine triphosphate with high purity of 97.2% and recovery of 82.5% was obtained. Compared with other conventional methods, the present separation process was simple and effective, and could be used in separation of nucleotides from microbial fermentation or transformation feedstocks.

Key words: cryogel, ion exchange, adsorption, separation, cytidine triphosphate

摘要： 用结晶致孔法制备了内嵌SiO₂颗粒的聚甲基丙烯酸羟乙酯基复合晶胶，经甲基丙烯酸二甲氨乙酯接枝修饰后，得到了阴离子交换复合晶胶介质，用于从啤酒酵母转化液中色谱分离三磷酸胞苷。色谱分离过程以0.01 mol·L^-1HCl为平衡液，以0.02 mol·L^-1 和0.1 mol·L^-1 的NaCl溶液（用平衡液配制）分步洗脱。通过高效液相色谱定量分析，所得三磷酸胞苷的纯度达97.2%，回收率82.5%。与其他分离方法相比，本法过程简单、分离迅速高效，在核苷酸药物的分离领域具有很好的应用前景。

关键词: 晶胶, 离子交换, 吸附, 分离, 三磷酸胞苷

CLC Number:

TQ028

LIU Jie, SHEN Shaochuan, CHEN Ping, XU Linhong, YUN Junxian, YAO Kejian. Separation of cytidine triphosphate from Saccharomyces cerevisiae broth by anion exchange poly(2-hydroxyethyl methacrylate) composite cryogel embedded with SiO₂ nanoparticles[J]. CIESC Journal, 2014, 65(10): 3938-3945.

刘杰, 沈绍传, 陈平, 徐林红, 贠军贤, 姚克俭. 内嵌纳米粒阴离子交换聚甲基丙烯酸羟乙酯复合晶胶分离三磷酸胞苷[J]. 化工学报, 2014, 65(10): 3938-3945.

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Separation of cytidine triphosphate from Saccharomyces cerevisiae broth by anion exchange poly(2-hydroxyethyl methacrylate) composite cryogel embedded with SiO₂ nanoparticles

内嵌纳米粒阴离子交换聚甲基丙烯酸羟乙酯复合晶胶分离三磷酸胞苷

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