小体系高通量比色测定糖化酶活力新方法

A new method of micro-system and high-throughput determination of glucoamylase activity by Colorimetry

本研究建立了一种小体系结合96孔板比色测定糖化酶活力的新方法。经实验证明,在96孔板内,波长540nm处,葡萄糖浓度与光密度（OD）值有很好的线性关系,其线性相关系数r值达到0.9996;通过加样回收实验发现,其回收率为98.5%~103.2%,相对标准差为1.8%;通过精密度分析实验发现该方法测量样品的精密度优于国标法及普通DNS法,其测定的结果与现行国家标准测定糖化酶方法所得到的结果差异率小于0.5%。本研究建立的方法不仅准确、可靠,而且实现了小体系、高通量快速比色测定,为糖化酶工业检测及糖化酶生产菌株的高通量筛选奠定了重要方法基础。

In this study,we reported a new assay method that a micro-system determined the activity of glucoamylase by dinitrosalicylic acid（DNS） reaction.In 96-well plates,glucose concentration and optical density（OD） values showed linear relationship at 540nm,and the linear correlation coefficient（r） values was 0.9996.The sample recovery experiment described that the recovery range and the relative standard deviation（RSD） was 98.5%~ 103.2% and 1.8%,respectively.The precision analysis to confirm the method is accurate and reliable.To compared with national standard method,the differences in rates of glucoamylase activity assay was less than 0.5%.The method is rapid,accurate and high-throughput on determination of glucoamylase activity,and will become an important method on quality control of commodity glucoamylase and screening of glucoamylase producing strain.