›› 2002, Vol. 45 ›› Issue (2): 209-214.

• 研究论文 • 上一篇    下一篇

二化螟体内乙酰胆碱酯酶的分布及纯化方法

彭宇, 王荫长, 韩召军, 陈长琨, 李国清   

  • 出版日期:2002-04-20 发布日期:2002-04-20

Distribution and purification of acetylcholinesterase in Chilo suppressalis

PENG Yu, WANG Yin-Chang, HAN Zhao-Jun, CHEN Chang-Kun, LI Guo-Qing   

  • Online:2002-04-20 Published:2002-04-20

摘要: 研究了二化螟Chilo suppressalis乙酰胆碱酯酶(AChE)的体躯和亚细胞分布,并用凝胶过滤层析和2种亲和层析方法从二化螟幼虫体内分离、纯化乙酰胆碱酯酶。结果表明:二化螟幼虫乙酰胆碱酯酶的活性主要集中于头部和胸部,而成虫胸部乙酰胆碱酯酶的活性最低,显著低于头部和腹部。成虫体内AChE的活性明显高于幼虫。在亚细胞的分布上,乙酰胆碱酯酶主要位于膜上(86%),近46%的活性存在微粒体中。在3种纯化乙酰胆碱酯酶的方法中,以3-羧基苯基-乙基二甲基铵作配体的亲和层析法纯化效果最佳,乙酰胆碱酯酶的最高纯化倍数为536.05倍,产率30.46%。

关键词: 二化螟, 乙酰胆碱酯酶, 亲和层析, 分布, 纯化

Abstract: The distribution in body and subcellular distributions and purification of cetylcholinesterase (AChE) in insecticide-resistant population of Chilo suppressalis were investigated. AChE was purified from the larvae of the same population by gel filtration and 2 species of affinity chromatography on 3-carboxyphenyl ethyldimethyl ammonium (CEA) and procainamide. The results showed that specific activity of AChE in adults was higher than that in larvae. AChE in larvae was distributed mostly in the head and the thorax; while in adults, AChE activity in thorax was lower than those in the head and the abdomen significantly. The major portion of the total AChE was member-bound (86%) and associated primarily with microsome (46%). Affinity chromatography on CEA was optimum for purification of AChE among 3 species purified methods. The extent of purification reached about 536.05-folds with recoveries of 30.46%.

Key words: Chilo suppressalis, acetylcholinesterase, affinity chromatography, distribution, purification