一种简便高效纯化重组D-氨基酸氧化酶的方法  

A Simple and High Efficient Method Used in Purifing Recombined D-Amino acid oxidase

作　　者：薛瀚 延国东 朱科[1] 王艳艳[1] 孟德程 

机构地区：[1]石药集团河北中润制药有限公司,河北石家庄050041

出　　处：《河北化工》2011年第10期18-20,共3页Hebei Chemical Industry

摘　　要：在重组D-氨基酸氧化酶上构建有6个组氨酸标签(6*Histag)的结构,用螯合了Ni2+的阳离子树脂FP-IDA可亲和吸附含6*Histag结构的蛋白,含DAO的酶溶液通过该层析柱时,被吸附在柱上。用低浓度咪唑溶液可洗脱杂酶,再用高浓度咪唑洗脱得到高纯度DAO溶液。用环氧型载体固定化DAO,经头孢菌素C裂解反应检测表明其中的杂酶活力极低,达到工业化要求。The recombined DAO was constructed a hexahistidine tag,and the Histagged DAO could be affinity adsorbed with a metal Ni2＋ chelate cation resin,DAO would be adsorbed on the chelate resin when the DAO solution flux to chromatography.The impurity enzyme could be eluted with low concentration of imidazole,then the DAO which was high purity could be eluted with the high concentration of imidazole.DAO was immobilized by epoxy carrier,then the immobilized DAO was test by the reaction of splitting cephalosporic C,the results suggested that the activity of impurity enzyme is very low,this product was fit to apply in producation.

关 键 词：D-氨基酸氧化酶 亲和吸附 头孢菌素C 

分 类 号：Q554.9[生物学—生物化学]