纳米磁粉固定化酶催化合成α-D-葡萄糖-磷酸.PDF

2010 Chinese Journal of Catalysis Vol. 31 No. 10 文章编号: 0253-9837(2010)10-1227-06 国际版DOI: 10.1016/S1872-2067(10)60116-7 研究论文: 1227~1232 纳米磁粉固定化酶催化合成 α-D-葡萄糖-1-磷酸 董 青, 欧阳立明, 刘建文, 许建和 华东理工大学生物反应器工程国家重点实验室, 上海 200237 摘要：建立了以麦芽糊精和磷酸盐为底物, 在常温下合成 α-D-葡萄糖-1-磷酸的生物催化体系. 从大肠杆菌 K12 中克隆表达了麦 芽糊精磷酸化酶, 并固定化在氨基修饰的磁性纳米颗粒上, 以便于酶的回收和重复利用. 在优化的反应条件下, 于 200 ml 体系中 连续使用该固定化酶 8 批次, 催化合成了 α-D-葡萄糖-1-磷酸. 经过简单的纯化步骤, 最终得到 440 mg 产品, 分离产率为 70.5%. 关键词：α-D-葡萄糖-1-磷酸; 麦芽糊精磷酸化酶; 麦芽糊精; 磁性纳米颗粒; 固定化生物催化剂 中图分类号：O643/Q55 文献标识码：A Efficient Synthesis of α-D-Glucose-1-Phosphate by Maltodextrin Phosphorylase Immobilized on Amino-functionalized Magnetic Nanoparticles * DONG Qing, OUYANG Liming, LIU Jianwen, XU Jianhe State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China Abstract: α-D-Glucose-1-phosphate (Glc-1-P) is an expensive intermediate in the biosynthesis of nucleotide glucose. This paper describes a biocatalytic system for the efficient synthesis of Glc-1-P from the low cost raw materials: maltodextrin and phosphate at ordinary tempera- tures. After molecular cloning and the expression of a maltodextrin phosphorylase (MalPase) gene from E. coli (Escherichia coli) K12, the resultant recombinant enzyme was immobilized on amino-functionalized magnetic nanoparticles for recycling and repeated use. Conditions for the biocatalytic reaction were optimized and the immobilized MalPase could be easily recovered and reused over eight cycles