啤酒酵母泥中一种新型SOD的提取.pdfVIP

第46卷第3期 应　 用　 化　 工 Vol.46 No.3 2017年3月 Applied Chemical Industry Mar.2017 啤酒酵母泥中一种新型SOD 的提取 李继伟,杨杰,张权威,张宇,邓祥宜 (武汉设计工程学院食品与生物科技学院,湖北 武汉　 430205) 摘　 要:啤酒酵母泥以异丙醇破壁处理,以磷酸钾缓冲液提取超氧化物歧化酶(SOD),通过单因素和正交实验优化 了提取条件。 结果表明,提取SOD的最佳条件是:以100%浓度异丙醇破壁处理150 min,以pH5.0磷酸钾缓冲液 为提取剂,液料比36 mL/ g。 在此条件下,SOD的得率为1257.2 U/ g。 纯化后的SOD酶液主要含有分子量大小为 25.46 kD和49.53 kD两种蛋白,其中分子量25.46 KD的蛋白与NCBI蛋白质数据库中baker’s yeast 中SOD 的分 子量相符合,该酶理论等电点为8.49,与中文文献中报道的SOD均不同,是一种新型SOD。 关键词:废弃啤酒酵母泥;超氧化物歧化酶(SOD);提取条件优化;生物信息学分析 中图分类号:TQ461;TQ464.9　 　 文献标识码:A　 　 文章编号:1671-3206(2017)03-0477-04 The extraction of a novel SOD from waste beer yeast mud LI Ji-wei,YANGJie,ZHANG Quan-wei,ZHANG Yu,DENGXiang-yi (College of Food Life Science and Technology,Wuhan Institute of Design and Sciences,Wuhan430205,China) Abstract:Beer yeast mud were treatmented by isopropyl alcohol to break thewall,using potassium phos- phate buffer to extract superoxide dismutase.The superoxide dismutase (SOD)extraction conditionswere optimized throughsingle-factortestandorthogonaltest.Theresultsshowedthattheoptimalextractioncon- ditionswere:breaking cell wall with 100% isopropyl alcohol for 150 min,using pH5.0 potassium phos- phate buffer as the extraction agent,and liquid-powder ratio36 mL/ g,under the condition,the extraction rate of SOD was 1 257.2 U/ g.The purified SOD solution contained two main proteins with molecular weight of25.46 kD and49.53 kD.The 25.46 kD protein matched the SOD of baker’s yeast in NCBI protein databaseverywell,andthetheoretical isoelectricpoint (pI)of theenzymewas8.49.It isanovel SODwhich is different to those published in Chinese references. Key words:waste beer yeast mud;superoxide dismutase (SOD);optimization of extraction condi