In this study, a novel method of continuous coupling of immobilized enzymatic hydrolysis reactor and membrane separation (CIEH-MS) was used to isolate the immunomodulatory peptides from type II collagen (CII) in chick sternal cartilage. The immobilized neutral protease was successfully prepared with an activity of 400.5 U/g. The CIEH-MS system loaded with immobilized neutral protease had high operational stability with enzyme decay constant of 0.0077 and half-life of 89.61 hr. Using a CIEH-MS system, the immunomodulatory peptides were obtained with lymphocyte proliferation of 66.23%, peptide yield of 23.43%, degree of hydrolysis (DH) of 22.41%, and permeate flux of 6.17 L/m2 h. The peptide fractions were further purified and the P3-2-4 fraction (RGQLGPM) with 760.4 Da molecular weights exhibited the highest lymphocyte proliferation activity (85.54%) and binding ability to human leukocyte antigen-DRB1 (HLA-DRB1) molecules (133.2 ng/ml). The results demonstrated that CIEH-MS system is an effective way to obtain immunomodulatory peptides from CII. PRACTICAL APPLICATIONS: Chick sternal cartilage is one of the by-products of meat processing, and it is often discarded as waste or used for low-value purposes. CII is the most abundant collagen in chick sternal cartilage, and recently studies have demonstrated that CII peptides possess the ability to induce immunologic tolerance for the treatment of chronic disease. In order to find potential applications for this by-product, immunomodulatory peptides from CII hydrolysates in chick sternal cartilage were isolated using a novel CIEH-MS system. The result showed that CII peptides exhibited a high immunomodulatory activity, and had a potential application in functional foods and medical fields.
Keywords: coupling of immobilized enzymatic hydrolysis reactor and membrane separation; immobilized neutral protease; immunomodulatory peptides; preparation; stability; type II collagen.
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