NIPAB法测定青霉素G酰化酶酶活力的分析和改进

Evaluation and Improvement of Determination of Penicillin G Acylase Activity using NIPAB Method

原NIPAB(2-硝基-5-苯乙酰胺基苯甲酸)法以乙醇为终止剂,采用该法测定发酵制得的青霉素G酰化酶酶活力时,常遇到终止反应不完全、杂质等非酶反应因素显著影响测定结果的问题,因此必须对发酵样品进行预处理。本研究基于反应产物吸光度变化速率,改进为动态方法测定酶活力,不需预处理和终止剂,提高了精确度并简化了操作。

Ethanol was usually used as the terminator of NIPAB （2-nitro-5-phenylacetamidobenzoic acid） method for the determination of penicillin G acylase activity. It was found that it couldn＇t stop the reaction thoroughly and the non-enzymatic interferences affected the result obviously so that the pretreatment of sample was essential. A kinetic method based on the absorbance-change rate of the product was established, in which sample pretreatment could be omitted resulting in the simplification of the process and the improvement of the accuracy of determination.