Method for analyzing sequences of absorbable whey peptides by using Caco-2 cells

Abstract

The invention discloses a method for analyzing sequences of intestinal absorbable whey peptides by using Caco-2 cells. The method comprises the following steps: carrying out enzymatic hydrolysis on whey protein by respectively using proteases with specific enzyme digestion sites, including alkaline protease, trypsin and chymotrypsin, carrying out ultrafiltration and Sephadex G-10 gel filtration chromatographic separation on hydrolysate to obtain whey peptides with different enzyme digestion sites, molecular weights, hydrophilic and hydrophobic properties and electrostatic charges; and after establishing and verifying Caco-2 cells, carrying out a whey peptide intestinal transport experiment by taking the Caco-2 cells as an intestinal absorption model, collecting the whey peptides transported from basal surface (BL) sides of the Caco-2 cells, analyzing and identifying amino acid sequences of transported whey peptides by virtue of an ultra-performance liquid chromatography-mass spectrometer (UPLC-MS), and analyzing the sequence features of the transported whey peptides which can be completely absorbed by small intestines. After the understanding of the sequences of the whey peptides which can be completely absorbed by the small intestines, polypeptides with relatively good oral absorption utilization degrees can be screened, and the field of functional foods and new drug development is expanded.

Description

technical field [0001] The invention relates to the establishment of a polypeptide sequence analysis method, especially the establishment of a intestinal absorbable whey peptide sequence analysis method. Background technique [0002] Whey protein is a by-product in the production process of cheese and casein, and its content accounts for 18% to 20% of the protein content in milk. The preparation of biologically active peptides by hydrolyzing whey protein with protease is a direction of whey development at present. Bioactive peptides are a general term for a class of physiologically active oligopeptides. It has been found that bioactive peptides derived from whey protein include blood pressure-lowering peptides, mineral element absorption-promoting peptides, and morphine peptides. These bioactive peptides can exert physiological regulation activity after being absorbed into the human body. Therefore, bioactive peptides derived from milk protein have great development and app...

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