磷酸激酶亲和色谱介质的制备_易喻.pdf

290 Chinese Journal of Biochemical Pharmaceutics 2006 27 5 易 喻, 王玉姣, 应国清 ( 浙江工业大学 药学院, 浙江 杭州310014) : 制备适合分离纯化磷酸激酶的亲和色谱介质 以纸纤维 载体, 环氧氯丙烷等 活化剂, 考察接臂配基对磷酸激酶吸附效果的影响, 确定亲和色谱介质制备的工艺路线, 并对主要工艺参数进行优化 己糖激酶纯化倍数可达20 倍以上, 酶活回收率近60% 以自制的亲和色谱介质分离纯化啤酒酵母中的 磷酸激酶, 成本低, 操作简单, 分离效果良好 : 磷酸激酶; 亲和色谱; 纯化; 己糖激酶; 配基 : O658 : A : 1005- 1678( 2006) 05-0290-04 Affinity chromatography materials for kinase YI Yu, WANG Yu-jiao, YING Guo-qin ( School fo Pharmacy , Zhej iang University of Technology , H angz hou 310014, China) Abstract: Purpose To synthesize the affinity chromato raphy medium to purify kinase. Methods The supporter paper fiber was activatedwith epoxy chloropropane etc. Accordin to the influence of spacer and dif- ferent li and on absorption of kinase, process route was optimized. Results More than 20-fold purified Hexoki- nase was obtained in one step with a yield of 60% . Conclusion This method is suitable for to kinase purifica- tion owin to its low cost and simple operation. Key words: kinase; affinity chromato rap; purification; hexokinase; li and ATP 2. 1 粗酶液的获取 , ( EC2. 1. 1. 7, HK) 2. 2 亲和色谱介质的制备 1, 6-( FDP) [ 1] 2. 2. 1 , , 30~ 35 h, , [ 2] FDP ; HK ; 0. 1 mol/ L NaOH , 2~ 3 h, pH + [3-4] K 7, 1 2. 2. 2 [ 5] , ATP-Na , ; 1[ 6] [ 7] 2 , ; 2. 3 蛋白质含量的测定