不溶性草酸氧化酶的包埋及草酸快速检测方法研究  

Study on the methods of embedding insoluble oxalate oxidase and rapid determination of oxalic acid

作　　者：胡一鸿[1] 张梦成[1] 许春会[1] 

机构地区：[1]湖南人文科技学院生命科学系,湖南娄底417000

出　　处：《广东农业科学》2011年第9期107-109,共3页Guangdong Agricultural Sciences

基　　金：湖南省教育厅重点项目(10A063);湖南省科技计划项目(2009FJ3159);湖南省大学生研究性学习和创新性实验计划项目(2010373)

摘　　要：通过研究不溶性草酸氧化酶包埋方法和草酸快速检测方法,建立氧电极法测定草酸的便捷体系。以大麦麸皮为原料制备不溶性草酸氧化酶,采用5%的琼脂糖包埋,检测包埋酶的稳定性和抗离子干扰能力;以包埋酶为工具酶,通过氧电极法检测草酸含量。试验结果表明:包埋酶连续反应2 h酶活性下降低于5%,贮存30 d活性下降低于8%;草酸检测线性范围0.1～0.8mmol/L,外标法草酸回收率为83%～97%。用氧电极法与显色法测定小麦草酸含量,两种方法的测定值无显著性差异。但氧电极法测定耗时短,反应体系简单,易于推广应用。The methods of embedding insoluble oxalate oxidase and the rapid determination of oxalic acid were studied in order to establish a convenient oxalic acid assay system with oxygen electrodes.Insoluble oxalate oxidase was prepared from barley bran and embedded by 5% agarose gel.Its stability and resistance to ion disruption were determined.Oxalic acid content was also determined with oxygen electrodes method using the embedded enzyme.The results showed that the activity of the embedded enzyme decreased less than 5% after 2 h consecutive reaction,and the activity of this enzyme decreased less than 8% after 30 d storage;the signals of oxalic acid concentration were linear with 0.1～0.8 mmol/L;recovery of oxalic acid using external standard was 83%～97%.Compared with colorimetric method,no significant difference was observed between these two methods in determining wheat oxalic acid.This method is less time-consuming,the reaction system is simple,and easy to perform.

关 键 词：包埋 不溶性草酸氧化酶 大麦麸皮 氧电极 琼脂糖 

分 类 号：Q814.9[生物学—生物工程]