【摘要】 采用溶胶-凝胶法,以丙烯酰胺作为单体,制备了基于有机-硅胶杂化整体柱的β-葡萄糖醛酸酶反应器。优化了硅酸甲酯和γ-(甲基丙烯酰氧)丙基三甲氧基硅的物质的量的比、丙烯酰胺和聚乙二醇的用量,以及水浴温度等制备条件,获得了孔隙均匀、通透性良好、机械强度高的有机-硅胶杂化整体柱。采用光学显微镜和扫描电镜表征杂化整体柱。进一步将β-葡萄糖醛酸酶共价键合在整体柱上,以4-甲基亚硝胺基-1-(3-吡啶)-1-丁醇(NNAL)的O-糖苷化合物(NNAL-O-Gluc)为底物研究酶反应器的水解效果,实验结果证明酶反应器在室温条件下的水解效率大大提高,实现了NNAL-O-Gluc高效水解与分析,解决了目前NNAL-O-Gluc分析中前处理水解效率低的问题。更多还原

【Abstract】 An immobilized β-glucuronidase enzyme reactor(IMER)based on organic-silica gel hybridized capillary column was prepared by the sol-gel method with acrylamide(AAM)as the organic functional monomer. The effects of the amounts of tetramethyl orthosilicate(TMOS),3-glycidoxypropyltrimethoxysilane(γ-MAPS),acrylamide(AAM),polyethylene glycol(PEG),and the water bath temperature on the quality of the microscopic structure were studied. After optimization,the monolithic column with good permeability and high mechanical strength was obtained. Then β-glucuronidase was covalently immobilized,and the hydrolysis efficiency of theβ-glucuronidase IMER was investigated. As the substrate,O-glucuronides of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol(NNAL-O-Gluc) were hydrolyzed at room temperature,indicating the high hydrolysis efficiency of the β-glucuronidase IMER,which demonstrated its high potential for the high throughput analysis of tobacco metabolites in the future.更多还原