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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。
OBJECTIVE: To study the effects of protective agent, solvent-dissolving agent and activator on the separation and extraction of superoxide dismutase (SOD) from porcine and bovine blood clots, and to determine its type and dosage. Methods: Two factors, three factors experimental design. Results: The difference of SOD activity and yield of pig blood and bovine blood reached the extremely significant level (P <0.01). The mixture of protective agent was better than single use, and the combination of 1.5% lactose (a) + 0.5% β-cyclodextrin (B) + 0.25% polyethylene glycol (c) had the best effect. The solvent-dissolving agent was 3% Triton X-100 (d) + 0.5% n-butanol % Triton X-100 (d) better effect; protective agent dosage to the clot quality of 20% is best, the amount of solvent-based clot quality 15% ~ 20% is better, the amount of activator to 2.5% best. CONCLUSION: It is unnecessary to add blood anticoagulant and centrifugation blood cell technology by blood sampling, and it has good effect and low cost by dissolving solvent and mechanical hemolysis, which is suitable for large-scale production. The SOD activity obtained by this process was above 3000 U / mg protein, and the activity and yield of SOD extracted from bovine blood were higher than that of pig blood.