酶解制备高溶解性蛋清蛋白粉工艺优化及其免疫调节活性研究

Optimization of Enzymolysis Parameters of Highly Soluble Egg White Protein Powder and Its Application in Immunoregulation in Vitro

鸡蛋清中的蛋白质种类丰富、生物效价高且具有多种生物活性,为充分利用高品质蛋清蛋白质资源,本实验在单因素实验的基础上选取酶解时间、酶解温度、酶解p H和加酶量四个因素进行四因素四水平的正交试验,以蛋清蛋白粉的溶解度和分散性为评价指标,优化了酶解工艺。实验得出用中性蛋白酶和木瓜蛋白酶以1:2的比例,加酶量800 U/g,酶解温度53.5℃,酶解p H 5.5,酶解时间90 min时,能使溶解度达到96.61%,分散时间达到39.62 s。同时通过细胞实验,得出高溶解性蛋清粉模拟胃肠道消化后作用于RAW264.7细胞,细胞内酸性磷酸酶、溶菌酶含量显著增多,且RAW264.7巨噬细胞增殖指数达到1.28,并显著提高了细胞因子等的分泌,具有免疫调节活性。该研究为开发溶解性好、品质高、功能效果明显的禽蛋蛋白质类产品提供了重要参考。

Egg white is rich in proteins and has high biological values and biological activities, which has been served as an excellent food resource. However, the egg white protein products have not yet been fully developed. Here, orthogonal experiment was designed by selecting four factors, including hydrolysis time, hydrolysis temperature, p H and enzyme dosage based on single factor experiments. The solubility and dispersibility of egg white protein powder were traced as evaluation indicators for optimizing enzymatic process. The results showed that the optimal solubility and dispersion time of egg white protein powder reached 96.6% and 39.6 s, respectively when the ratio of neutral protease to papain, the enzyme amount, the hydrolysis temperature, enzymatic hydrolysis time and p H being of 1：2, 800 U/g, 53.5 ℃, 90 min and 5.5, respectively. Furthermore, the effects of simulate gastrointestinal digestion products（SGDP） derived from highly soluble egg white protein powder on the RAW264.7 macrophages was studied by cell experiment. The results showed that SGDP significantly increased the level of acid phosphatase（ACP） and lysozyme（LZM） intracellular. Meanwhile, the role of SGDP in immunoregulation was confirmed by the enhanced proliferation potential（proliferation index 1.28） and cytokine release. This study can provide an important reference for the development of egg protein products with highly soluble and function.