蛹虫草菌丝体中SOD提纯工艺及鉴定  

Purification and Isozyme Identification of SOD from Cordycepsmilitaris Mycelia by Submerge Fermentation

作　　者：杜琳[1] 邱芳萍[1] 邢燕[1] 纪可[1] 

机构地区：[1]长春工业大学化学与生命科学学院,吉林长春130012

出　　处：《食用菌学报》2013年第4期49-54,共6页Acta Edulis Fungi

基　　金：长白山生态资源综合利用与开的平台建设(编号:721002)的部分研究内容

摘　　要：深层发酵蛹虫草菌丝体中饱和硫酸铵分级沉淀法提取超氧化物歧化酶（SOD）,单因素实验基础上,运用响应面法优化提取工艺,获得最佳SOD提取工艺参数,在此条件下将粗酶液上Sephadex G-100柱和DEAESepharose FF柱纯化,并采用紫外光谱扫描、SDS-PAGE法、原子吸收分光光度法对SOD分子结构上所携带的金属辅基进行了鉴定。结果表明：SOD的最佳提取工艺条件：液料比22∶1、pH8.2、提取时间1.7h,总酶活力为142.1U/mL,纯化倍数为3.78;全波长扫描在258nm和265nm有吸收峰,SOD相对分子质量在25 000~35 000之间,Cu、Zn的含量分别为2.06%、3.88%。因此,深层发酵蛹虫草菌丝体中SOD类型为Cu/Zn-SOD。SOD（superoxide dismutase）was extracted with phosphate buffered saline（PBS）from dried Cordyceps militaris mycelium grown in submerged culture.After centrifugation（4460 g,15 min）,the enzyme precipitating between 10%-100% ammonium sulfate was collected.On the basis of single factor experiments,response surface methodology was used to optimize the SOD extraction conditions,which were：20∶1ratio of PBS buffer to dried fungal mycelium powder,pH 8and an extraction time of 1.5h.Under these conditions,142U/mL of SOD was obtained.The crude extract was purified by Sephadex G-100and DEAE-Sepharose FF column chromatography,and the purification fold was 3.78.UV scanning spectroscopy revealed that purified SOD had absorption peaks at 258nm and 265nm,and SDS-PAGE indicated the molecular weight of the enzyme to be between 25 000and 35 000.Cu and Zn contents,determined by atomic absorption spectrophotometry,were 2.06% and 3.88%,respectively.

关 键 词：蛹虫草 菌丝体 SOD 

分 类 号：S567.35[农业科学—中草药栽培]