猪脾脏多肽的酶法制备及其生物活性研究.pdfVIP

食品与药品 Food and Drug 2010年第 12卷第05期 161 猪脾脏多肽的酶法制备及其生物活性研究 * 刘庆涛，余 蓉 ，崔瑜霞，马宇峰，吴城光，杨继虞 （四川大学 华西药学院生物制药教研室，四川 成都 610041 ） 摘 要：目的 探索酶解法制备猪脾脏多肽的最佳条件，检测所得产物的生物活性。方法 用胰酶酶解法、正交试验、甲醛 滴定法考察最佳酶解条件，用乙醇沉淀法收集多肽，并分别用Fe2+诱发卵黄脂蛋白多不饱和脂肪酸过氧化体系和纤维蛋白 平板法检测多肽的抗氧化活性和溶栓活性，CM-Sepharose离子交换层析纯化最佳酶解条件下的脾多肽，利用SDS电 泳确定具备促凝活性的多肽分子片段相对分子质量。结果 最佳酶解条件为60 ℃，pH 7.0，加酶量6.0 %，时间5.5 h ；检测 到脾多肽具备抗氧化活性和促凝活性，经纯化后的脾多肽促凝时间比正常对照提前20 s 。 结论 可初步判断猪脾脏多肽在 抗氧化和凝血方面有一定作用，且首次证实猪脾多肽具有凝血功能。 关键词：猪脾脏；多肽；抗氧化；促凝 中图分类号：Q556 文献标识码：A 文章编号：1672-979X(2010)05-0161-04 Preparation of Pig Spleen Peptides by Enzymatic Hydrolysis and Its Biological Activity LIU Qing-tao, YU Rong, CUI Yu-xia, MA Yu-feng, WU Cheng-guang, YANG Ji-yu ( Division of Biop harmaceutics, Pharmacy School of West China, Sichuan University, Chengdu 61004 1, China) Abstract: Objective To explore the best conditions of preparation of pig spleen peptides by enzymatic hydrolysis， and to detect the biological activity of enzyme reaction products. Methods Trypsin digestion method, orthogonal experiment, and formaldehyde titration method were used to study the optimal hydrolysis conditions. The peptides were collected by ethanol precipitation, then Fe2+ was used to induce PUFA peroxidation system in egg yolk lipoprotein. The fibrin plate method was employed to assay the antioxidant activity and thrombolytic activity of peptides . The spleen polypeptides obtained under the optimal hydrolysis conditions were purified by CM-Sepharose ion-exchange chromatography and the molecular weight of the peptide fragments with procoagulant activity was determined by SDSelectrophoresis. Results The optimal enzymatic hydrolysis conditions of pig spleen peptides were 60 ℃, pH 7.0, 6.0 % e