重组精氨酸脱亚胺酶的酶活性测定

Enzyme Activity Determination of Recombinant Arginine Deiminase

建立了大肠杆菌表达的重组精氨酸脱亚胺酶（1）的酶活性体外测定方法。通过优化反应体积、底物浓度、反应时间、反应液pH值等参数,确定测定重组1酶活性的最佳条件为：底物精氨酸浓度20 mmol/L、pH 6.5、反应时间20 min、反应体积100ul（试管法）。本法的线性范围为10～25ug/ml,变异系数（CV）小于10%。

An assay method was established for the enzyme activity determination of recombinant arginine deiminase （ADI） expressed by E. coli. The effects of reaction solution volume, substrate concentration, reaction time and pH value of reaction solution on enzyme activity were investigated. The optimized conditions were as follows： 20 mmol/L arginine, pH 6.5, reaction time of 20 min and reaction volume of 100 μl （test tube method）. The calibration curve of ADI was linear in the range of 10 - 25 μg/ml with CV less than 10%.