响应面法优化燕窝不溶性蛋白酶解工艺的研究

The Response Surface Methodology Optimizes Enzymatic Hydrolysis Process of Insoluble Protein in Edible Bird's Nest

研究旨在应用酶解技术将燕窝中不溶性蛋白转换为水溶性多肽,有利于人体消化吸收,以期为燕窝新产品的开发奠定基础。以水解度和肽得率为指标,筛选出碱性蛋白酶对燕窝中不溶性蛋白进行酶解。通过单因素实验研究酶解时间、酶加量、底物浓度对燕窝多肽得率的影响,并利用响应曲面法对燕窝不溶性蛋白酶解工艺进行优化。酶解的最佳工艺为:底物浓度为2.5%,酶加量为4%,酶解时间为3.6 h,肽得率为33.59%。通过试验对该模型进行验证:得到肽得率为33.38%,相对误差为0.56%,说明该模型能较好地反映出燕窝不溶性蛋白酶解情况。此外,通过凝胶电泳试验证明燕窝不溶性蛋白通过碱性蛋白酶酶解,分子量有效降低。该试验获得了燕窝多肽,为后续产品进一步深入开发和应用提供一定理论基础。

The research aims to use the enzymolysis technology to convert the insoluble protein from edible bird’s nest into water-soluble peptide, which could facilitate the digestion and absorption by the human body,and provide a foundation for the development of new products of edible bird’s nest. Using the hydrolyzing degree and the yield of peptide as index, alkaline protease was selected to hydrolyze the insoluble protein from edible bird’s nest. The effects of enzymolysis time, enzyme amount and substrate concentration on the yield of edible bird’s nest peptide were studied with single factor experiment. The response surface analysis was used to optimize the enzymatic conditions of the insoluble protein from edible bird’s nest. The optimal condition of enzymatic hydrolysis was: substrate concentration of 2.5%, the enzyme amount of 4%, enzymolysis time of 3.6 h, and the yield of peptide of 33.59%. The model was verified by the experiment: the yield of the peptide was33.38%, and the relative error was 0.56%, which indicated that the model could well reflect the enzymatic hydrolysis of the insoluble protein from edible bird’s nest. In addition, gel electrophoresis experiments showed that the molecular mass of the insoluble protein was effectively reduced by using alkaline protease to hydrolyze the insoluble protein.