碱性蛋白酶酶解蛋白制备活性肽的质控研究

Quality Control of Active Peptides Preparation Through Hydrolyzing Protein by Alkaline Protease

以胶原蛋白和大豆分离蛋白为主要研究对象,应用体积排阻色谱(SEC)结合多角度激光光散射(MALLS)及示差折光(RI)联用技术对碱性蛋白酶酶解蛋白制备肽段的过程进行了详细分析。在酶解过程中,不同来源的胶原蛋白(牛、鸡)和大豆分离蛋白在分子量分布、体系的分散度及粒径变化规律上表现出了典型的差异。结果表明,牛胶原蛋白在酶解开始时就迅速被降解产生了大量小分子肽段,鸡胶原蛋白相对于牛胶原蛋白在相同的酶解条件下更难产生小分子肽段,大豆分离蛋白随着酶的加入则是表现出了先聚集再降解的特性。SEC-MALLS技术可以有效把控蛋白酶解的过程以及体系中肽段分子量分布,该技术可以为活性肽产品的开发及品质检测提供依据。

The main research objects were collagen and soybean protein isolate,and the process of preparing pep tide segments through hydrolyzing protein by alkaline protease was analyzed in detail by using size exclusion chromato graphy(SEC)combined with multi-angle laser light scattering(MALLS)and refraction index(RI).During the enzy molysis process,collagen from different sources(bovine and chicken)and soybean protein isolate showed typical differences in change rules of molecular weight distribution,dispersity and root-mean-square radius.The results showed that bovine collagen was rapidly degraded at the beginning of enzymolysis and a large number of small molecule peptides were produced.Chicken collagen was more difficult to break down to produce smaller peptides than bovine collagen under the same enzymolysis conditions.With the addition of enzyme,soybean protein isolate showed the characteristics of first aggregation and then degradation.SEC-MALLS technology can effectively control the process of proteolysis and the molecular weight distribution of peptides in the system.This technique can provide a basis for the development and quality testing of active peptide products.