【作者】 郭书贤； 王冬梅； 薛刚； 刘凤霞； 高伟霞；

【Author】 Guo Shuxian Wang Dongmei Xue Gang Liu Fengxia Gao Weixia (School of Biochemical Engineering,Nanyang institute of technology,Nanyang 473004)

【机构】 南阳理工学院生物与化学工程学院；

【摘要】 目的：研究保护剂、溶膜剂和激活剂对从猪、牛血块中分离提取超氧化物歧化酶 (SOD)的影响,确定其种类和用量。方法：采用两因素、三因素试验设计。结果：猪血与牛血在 SOD 活性和产率上差异达到极显著水平(P＜0．01)；保护剂混合组合好于单用,混合组合以1．5%乳糖(a)+0．5%β-环糊精(b)+0．25%聚乙二醇(c)效果最好；溶膜剂：牛血以3%曲拉通 X-100(d)+0．5%正丁醇(e),猪血以3%曲拉通 X-100(d)效果较好；保护剂加量以血块质量的20%为最佳,溶膜剂加量以血块质量的15%～20%为好,激活剂加量以2．5%最佳。结论：省去了采血加入抗凝剂和离心分离血球工艺,且采用溶膜剂及机械法溶血,效果好、成本较低,适合规模化生产。以此工艺获得的 SOD 活性在3000U/mg 蛋白以上,且从牛血提取的 SOD 活性和产率均高于猪血。更多还原

【Abstract】 Objective:To study the effect of protective agent,membrane lytic agent and activator on SOD extracted from pig and bovine blood block.Method:Double and three factor test designs were adopted.Results:The gap of active and productive rates of SOD between pig and bovine blood block reached very obvious level(P＜0.01);The mixture protective agent was better than single,the best was 1.5% galactosylglucose(a)+0.5% β-cyclodextrin(b)+0.25% PEG(c), and the better membrane lytic agent was 3% Triton X-100(d)+ 0.5% n-butanol(e)for bovine blood,and 3% Triton X- 100(d)for pig blood.The optimal dosage of protective agent was 20% of blood block weight,the optimal dosage of membrane lyric agent was 15%～20% of blood block weight,and the best optimal dosage of activator was 2.5%.Conclusion:This technique not only omitted the anticoagulant and centrifugation for separation of blood cell,but also effectively adopts membrane lyric agent and mechanism method for haemolysis at lower cost and convenience to fit mass production.As a result,the higher purity SOD was obtained,whose activity exceeded 3 000 U/mg protein,the activity and productive rates of SOD extracted from bovine blood block were higher than pig blood block.更多还原