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 fromLocusta migratoria and preliminary study on pesticide screening

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SDSPAGE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Ⅱ3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)%.

AChE was purified by affinity chromatography from Locusta migratoria head. The purifica- tion 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 cyto- toxicity and inhibitory effects was discussed. The results showed that 19 compounds had inhibitory effects on AChE and 13 compounds with cytotoxicity on AWl cells. Compound Ⅱ had shown better rel- evance to AChE activity with cytotoxicity . The AChE inhibition rate of Ⅱ_3 was （80.94 ＋ 3.09） % un- der the concentration of 100 mg/L and cytotoxicity activities was （95.01±2.24） % under the concentra- tion of 50 mg/L.