食品科学 ›› 2012, Vol. 33 ›› Issue (8): 6-10.doi: 10.7506/spkx1002-6630-201208002

• 工艺技术 • 上一篇    下一篇

海参肠乙酰胆碱酯酶亲和层析洗脱条件的优化

杜 英,朱蓓薇,吴海涛*   

  1. 大连工业大学食品学院
  • 出版日期:2012-04-25 发布日期:2012-03-31
  • 基金资助:
    国家自然科学基金青年科学基金项目(31000754);辽宁省博士科研启动基金项目(20101004)

Optimization of Elution Conditions for Affinity Chromatography of Acetylcholinesterase from the Gut of Sea Cucumber (Stichopus japonicus)

DU Ying,ZHU Bei-wei,WU Hai-tao*   

  1. (School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, China)
  • Online:2012-04-25 Published:2012-03-31

摘要: 以3-羧基苯基-乙基二甲基铵为配基,与溴化氰活化琼脂糖凝胶交联制备亲和层析柱,分离海参肠中乙酰胆碱酯酶(acetylcholinesterase,AChE),对其洗脱条件进行优化。结果显示:其适宜平衡体系为含0.3mol/L NaCl的PBS(0.05mol/L,pH7.4)缓冲溶液,适宜洗脱体系为含0.2mol/L四乙基碘化铵的PBS(0.05mol/L,pH7.4)缓冲溶液。此条件下,海参肠AChE可得到较好的吸附和分离,初步纯化后的AChE经Native-PAGE电泳分离出3条具有酶活性的条带,说明AChE在海参肠内可能以多种形式存在,呈现多态性。

关键词: 海参, 乙酰胆碱酯酶, 亲和层析, 洗脱条件

Abstract: An affinity chromatography column was prepared by cross-linking of 3-carboxyphenyl ethyldimethyl ammonium (CEA) as a ligand with CNBr-Sepharose-4B and used for an investigation into the optimization of elution conditions for affinity chromatography of acetylcholinesterase (AChE) from the gut of sea cucumber (Stichopus japonica). The appropriate balance system was PBS (0.05 mol/L, pH 7.4) containing 0.3 mol/L NaCl, and the appropriate elution system was PBS (0.05 mol/L, pH 7.4) containing 0.2 mol/L tetraethyl ammonium iodide. Under these conditions, AChE was well adsorbed and separated. Three active bands were separated in the native-PAGE pattern of Preliminarily purified AChE, AChE in the gut of sea cucumber may have different forms and display polymorphism.

Key words: sea cucumber, acetylcholinesterase, affinity chromatography, elution conditions

中图分类号: