东亚飞蝗乙酰胆碱酯酶的分离纯化与农药活性筛选初步研究

Purification of Acetylcholinesterase from Locusta migratoria and preliminary study on pesticide screening

摘要:

采用亲和层析法对东亚飞蝗（Locusta migratoria ）脑组织的乙酰胆碱酯酶（Acetylcholinesterase，AChE）进行分离纯化，纯化倍数为474.81倍，产率为28.93 %，此酶经过SDSPAGE呈一条带，达到电泳纯，相对分子质量约为67.22kDa.同时测定了课题组前期合成的24种呋喃酮类似物对该乙酰胆碱酯酶的抑制活性和对RPHzVNCAW1（AW1）细胞的毒力，并分析其相关性.结果表明：在24种呋喃酮类似物中，对AChE有抑制作用的化合物有19种，对AW1细胞有增殖抑制作用的有13种.结果还显示，属于第Ⅱ类化合物的细胞毒力与AChE抑制活性关联性好于其它两类，其中Ⅱ3化合物，在100mg/L的浓度下，AChE抑制率为（80.94±3.09）%；在50mg/L的浓度下，对AW1的细胞毒力为（95.01±2.24）%.

Abstract:

AChE was purified by affinity chromatography from Locusta migratoria head. The purification factors and yields were 474.81fold and 28.93%, respectively. The molecular weight of the purified enzyme was 67.22kDa, measured by SDSPAGE. The inhibitory activity of 24 furanone analogs against AChE and the toxic effect against RPHzVNCAW1（AW1）were measured, then the relevance of cytotoxicity and inhibitory effects was discussed. The results showed that 19 compounds had inhibitory effects on AChE and 13 compounds with cytotoxicity on AW1 cells. Compound Ⅱ had shown better relevance to AChE activity with cytotoxicity . The AChE inhibition rate of Ⅱ3 was (80.94 + 3.09)% under the concentration of 100 mg/L and cytotoxicity activities was (95.01±2.24) % under the concentration of 50 mg/L.