关键词: 鹿茸肽, 碱性蛋白酶, 酶解, 抗氧化

Abstract: An industrial alkaline protease Alcalase was used to hydrolyze the fresh velvet antler of sika deer (Cervus Nippon Temminck). The conditions of enzymatic hydrolysis were optimized. The effects of several factors, including amount of the enzyme, substrate concentration and hydrolysis time, on the antioxidative activities of the hydrolysis product were examined by orthogonal test design. The optimum hydrolysis conditions were established to be the amount of enzyme, 1:150; the substrate concentration, 1:13; hydrolysis time, 60min. The crude velvet antlers peptides were prepared by fractionation of ammonium sulphate. An elution peak with high antioxidative activities from Sephadex G-25 column chromatography of the crude velvet antlers peptides was collected and named as VAP-B. Finally, a further purified velvet antlers peptide component (VAP-B1) with molecular weight distribution from 200-800 was obtained from VAP-B by preparative HPLC. The protein content of VAP-B1 was 70.45%. An analysis of amino acid composition was also carried out. The antioxidative activities of velvet antlers peptide component (VAP-B1), including removal of superoxide anions, hydroxyl radicals and inhibition of lipid peroxidation were examined. The reducing power of VAP-B1 was also tested. The experimental results indicate that when VAP-B1 concentration was 20 mg/ml, the inhibition ratio to pyrogallic acid autoxidation reached to 91.9%, having a strong ability to scavenge superoxide anion; when VAP-B1 concentration was 10 mg/ml, the clearance rate to hydroxyl radical was 100% and was dosage-dependent in a certain rage of concentration; At the same time, VAP-B1 showed some capability of inhibition of lipid peroxidation and reducing power.

Key words: Velvet antlers peptides, Alcalse, Enzymatic hydrolysis, Antioxidative activities