酶电极法快速测定甘油含量的研究  

Study of Rapid Determination for Glycerol Content by Enzyme Electrode

作　　者：司洪宇[1,2] 王丙莲[3,4] 梁晓辉[1,2] 张晓东[1,2] 

机构地区：[1]山东省科学院能源研究所,济南250014 [2]山东省生物质气化技术重点实验室,济南250014 [3]山东省科学院生物研究所,济南250014 [4]山东省生物传感器重点实验室,济南250014

出　　处：《中国生物工程杂志》2016年第12期79-85,共7页China Biotechnology

基　　金：山东省重点研发项目(2015GSF121047);山东省重点研发项目(2015GSF117034);公益性行业(农业)科研专项(201503135-04);山东省自然科学基金(ZR2016YL007)资助项目

摘　　要：利用酶固定化技术,以甘油激酶（GK）、甘油-3-磷酸氧化酶（GPO）为反应酶,研究GK、GPO的固定化方法及固定化模式,制备甘油酶膜、甘油酶电极,并利用其测定甘油含量。结果表明,GK、GPO按1∶1比例固定化时,酶电极电流信号最高;最高效固定模式为：GK固定于核微孔膜,共价偶联GPO固定于Biodyne膜,形成共价双酶膜,进而组装为甘油酶电极。性能研究表明,甘油酶电极最适pH值为7.0,最佳温度为28~32℃;最佳实验条件下,线性范围为0.05~9.00 g/L;回收率为98.4%~102.4%,稳定性高,相对标准偏差（RSD）〈5%;测定结果与高效液相色谱法、高碘酸氧化法比较,无明显差异（P〉0.05）,且该方法操作简单,专一性强,检测快速,适于实际生产中甘油的实时定量及监控。Glycerokinase（ GK） and glycerol-3-phosphate oxidase（ GPO） were immobilized to develop glycerol enzyme membrane and enzyme electrode for determination of glycerol concentration. Results showed that the higher current output for glycerol was obtained using a 1∶ 1（ GK∶ GPO） ratio for the random immobilization.The efficient two-enzymatic membrane was prepared by immobilizing GK in nuclear pore film coupled with GPO on Biodyne B. Glycerol enzyme electrodes were realized by assembling the GK-GPO membranes previously prepared. In this way,the optimum pH was 7. 0,and the fit range of temperature was 28 ～ 32℃. In the best experimental conditions,the linear scope was 0. 05 ～ 9. 00 g/L with excellent linearity and high stability. The recovery was 98. 4% ～ 102. 4%,and its relative standard deviation（ RSD） was〈 5%. There was no significant difference（ P〉0. 05） in measurement results among glycerol enzyme electrode,Periodate oxidation method and HPLC.

关 键 词：甘油 固定化酶膜 酶电极 

分 类 号：Q819[生物学—生物工程]