三嗪染料修饰曲通X-100亲和分离苹果酸脱氢酶
Chemical Research and Application(2011)
Abstract
三嗪染料(Cibacmn blue F3G-A)的蒽醌部分结构类似于腺嘌呤,可以用于亲和分离以NAD+(NADP+)和FAD为辅酶的脱氢酶.三嗪染料通过亲核取代反应修饰曲通X-100,形成的三嗪染料-曲通X-100与成相聚合物吐温80、磷酸钾盐构成液-固亲和萃取体系.从猪心肌匀浆液纯化苹果酸脱氢酶的条件:三嗪染料-曲通X-100浓度为1.2%(W/V),吐温舳浓度9%(V/V),猪心肌匀浆液0.25 mL(2.5%),pH8.0,加入成相磷酸钾终浓度1.6 mol/L,形成液固两相,固相酶活力平均收得率为81.01%,纯化倍数为4.93.固相30℃保温溶解,加入成相盐磷酸钾,终浓度为0.7mol/L,二次成相.低盐浓度条件下,近80%的酶反萃回液相,简化了苹果酸脱氢酶后续加工工艺.
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Key words
Triton X-100,malate dehydrogenase,Tween 80,Ctriazine-dye,liquid-solid affinity partitioning
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