【摘要】 对8种常用蛋白酶进行筛选,确定最佳用酶,并对酶解产物进行SDS-PAGE表征。再采用响应曲面试验方法(RSM)对胰蛋白酶水解工艺进行优化,利用Design-Expert7.1.6软件进行统计分析,建立回归模型。结果表明,以胰蛋白酶为水解最佳用酶,抑菌圈直径为14.62mm;SDS-PAGE表征可看出酶解物主要由小肽分子组成。胰蛋白酶水解最优工艺条件为:底物浓度([S])=11.84%,酶添加量([E])4.05‰,时间5.86h,pH8.5,温度40℃,在此条件下可获得抑菌圈直径达到16.54mm的水解产物。验证实验显示,建立的回归模型准确可靠。更多还原

【Abstract】 Enzymolysis was applied to prepare antibacterial peptides from Tenebrio molitor protein. Tenebrio molitor protein was hydrolyzed with eight proteases respectively and among them trypsin was found to have the best comprehensive performance confirmed by the antibacterial activity and SDS pattern of corresponding hydrolysates as well as peptide concentration in their solution. The trypsin-catalyzed hydrolysis of Tenebrio molitor protein was optimized to attain the highest antibacterial activity using response surface design and a regression model for antibacterial activity as a function of substrate concentration,enzyme dose and hydrolysis duration were built up. Under optimized conditions:substrate concentration 11.84%,enzyme dose 4.05‰,pH 8.5 and temperature 40 ℃ for a hydrolysis duration of 5.86 h,the actual diameter of antibacterial circle of hydrolysates reached 16.54 mm,close to the predicted value of 16.98 mm.更多还原