双酶水解黄粉虫蛋白制备生物活性肽的工艺优化

Study on preparation technique optimization of bioactive peptides from bienzyme-hydrolyzed Yellow mealworm protein by response surface method

【目的】对胰蛋白酶与碱性蛋白酶Alcalase双酶组合水解黄粉虫蛋白制备生物活性肽的工艺进行优化,为获得高活性黄粉虫蛋白多肽及有效利用黄粉虫蛋白提供科学依据。【方法】以水解度和酸溶性肽得率为指标,研究了酶解时间、酶活比、料液比、加酶量、pH和酶解温度6种因素对酶解反应的影响。在此基础上设计了3因素(加酶量、pH和酶解温度)3水平的响应面试验。【结果】胰蛋白酶与碱性蛋白酶Alcalase双酶水解黄粉虫蛋白的最佳酶解条件为:酶解时间120 min,酶活比1∶1,料液比1∶3,加酶量23.41 mg/g,pH 8.77,酶解温度53.41℃。【结论】利用优化双酶水解条件制得的低分子多肽的水解度高达21.61%,酸溶性肽得率为92.09%。

[Objective] In order to obtain the bioactive peptides,the Yellow mealworm protein was hydrolyzed by the mixture of Trypsin and Alkaline Protease and the conditions of enzymatic reaction were determined and optimized. [Method] By measuring the degree of hydrolysis （DH）and the yield of acid-soluble peptide （YASP）, time, the effects of ratio of protease energy, ratio of solid to liquid, dosage of protease, pH,and temperature on DH and YASP were studied in single factor test. On this basis,three factors （dosage of protease,pH,and temperature） and three levels＇ test by response surface method （RSM） were designed. [Result] The optimum conditions were：time 120 min,ratio of protease energy 1 ： 1,ratio of solid to liquid 1 ： 3,dosage of protease 23.41 mg/g,pH 8.77,and temperature 53.41 ℃. [Conclusion] Under these conditions,DH was 21.61% ,and YASP 92.09%.