胰蛋白酶水解全酪蛋白反应过程中的色谱分析

Chromatographic Analysis on Tryptic Hydrolysis of Whole Casein

将高效凝胶排阻色谱 (HPSEC)技术与水解度 (DH)概念相结合 ,对酪蛋白 胰蛋白酶水解体系的酶解反应过程进行色谱分析 ,得到定量表征复杂酶解反应进程和不同DH值时多样性酶解产物相对分子质量分布的二维图线 ;依据蛋白质结构信息 ,结合HPSEC实验谱图 ,对胰蛋白酶作用于酪蛋白时的酶解断裂位点进行剖析 ,初步推断反应历程 ,并得到理论酶解肽段的相对分子质量分布图及酶解物中活性多肽酪蛋白磷酸肽 (CPPs)肽谱。

Tryptic hydrolysis of whole casein was investigated by high performance size exclusion chromatography (HPSEC) in combination with the degree of hydrolysis (DH) In terms of HPSEC chromatograms obtained at different DH values, the complex process of enzymatic reaction and the relative molecular mass distribution of multiple hydrolysates were quantitatively characterized Based on the information of casein micelle structure, the possible reaction mechanism was deduced from a series of chromatograms Being taken into account the primary structure of whole casein and the target amino acid of trypsin, the distribution of theoretical peptides were accurately calculated by determining the split sites of complete enzymatic hydrolysis According to the relationship between retention time and relative molecular mass, the corresponding HPSEC absorption peaks of active peptides in hydrolysates were identified, and caseinophosphopeptides sequences were also characterized