A Study On Extracting Technology Of Collagen With Antler Base And ACE Inhibitory Activity

Deer antler base contains large amounts of collagen, which contains abundant ACE inhibitory activity of peptides, so as to collagen has important significance in the development of ACE inhibitory peptide raw materials, Seeking and finding the unique biological active substances from antler base polypeptide substance is the focus of this study.Take antler base of Cervus elaphus as material, after preprocessing, collagen was extracted from antler base of Cervus elaphus using pepsin and acetic acid. Extraction process for collagen of antler base was optimized by response surface methodology. The structure, molecular weight distribution and amino acid composition of collagen from antler base of Cervus elaphus were analyzed by ultraviolet spectra, infrared spectroscopy, SDS-PAGE and high performance liquid chromatography(HPLC) respectively. A comparative study was carried out to choose the best enzyme from pepsin, papain, neutral protease, trypsin and alkali protease for ACE inhibitory peptides preparation by enzymatic hydrolysis with collagen of antler base. ACE inhibitory peptides extraction process was optimized by response surface methodology. In this study, emulsification and emulsion stability, foaming and foam stability, hydroscopicity and moisture retention, oil absorption indexes of both collagen and ACE inhibitory peptides of antler base were studied. ExperimentⅤ: In this study, ACE inhibitory peptides of antler base were preliminary separated and purified by ultrafiltration, Sephadex G-25 and Sephadex LH-20. Experimental results are summarized as follows:1. Extraction process for collagen of antler base was optimized as p H, enzyme loading, hydrolysis temperature and hydrolysis time were 1.8, 4%, 37℃ and 5h by response surface methodology. Under the optimized conditions, collagen yield with antler base was 83.32%.2. ACE inhibitory peptides extraction process was optimized as p H, enzyme loading, hydrolysis temperature and hydrolysis time were 8, 5%, 55℃, 4h by response surface methodology. Under the optimized conditions, ACE inhibitory activity and DH could be reached to 82.07%、22.78% respectively.3. Emulsification of collagen of antler base and ACE inhibitory peptides were 70.4%, 40.2%; emulsion stability were 53.7%, 75.2%; foaming were 33.6%, 18.7%; foam stability were 76.2%, 50.4% respectively. Hydroscopicity increased with the relative humidity and time. Moisture retention decreased with the increase of relative humidity and time. Their oil absorption indexes were 1.2mg/m L, 2.1mg/m L respectively.4. After preliminary separation and purification, a higher ACE activity of fraction III-2-2 was obtained, its inhibitory rate was 97.72%, and its IC50 value was 0.02mg/m L. Compared with the separation and purification before, this inhibitory peptide ACE IC50 value was reduced by 2.8 times.Preparation of ACE inhibitory peptides by enzymolysis with the collagen of antler base provided both the theory basis for the research and development of ACE inhibitory peptide and the reference for deeply development and utilization of deer antler.