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乙酰胆碱酯酶(Acetylcholinesterase,AChE)是保证神经信号在生物体内正常传递的关键酶,是有机磷和氨基甲酸酯类农药的作用靶标.本研究在前期朱砂叶螨乙酰胆碱酯酶基因原核表达及最适反应条件摸索的基础上,大规模表达纯化朱砂叶螨AChE,进一步研究其酶学性质.研究结果显示,在温度为25℃、其他条件最适的情况下,测定酶K_m值为0.67 mmol/L,K_(cat)/K_m值为13.53 mmol L~(-1) s~(-1).随后,从SPECS化学库筛选到了55个候选AChE抑制性化合物,获得了9个抑制活性优于或相当于毒扁豆碱的朱砂叶螨AChE抑制化合物,并且它们对朱砂叶螨有较好的毒性.本研究为乙酰胆碱酯酶抑制剂的筛选以及以乙酰胆碱酯酶为靶标的农药开发提供了理论依据和实验基础.
Acetylcholinesterase (AChE) is the key enzyme to ensure the normal transmission of neural signals in vivo and is the target of organophosphorus and carbamate pesticides.In this study, the prokaryotic expression of acetylcholinesterase gene On the basis of exploring the reaction conditions, the AChE was purified and expressed on a large scale, and its enzymatic properties were further studied.The results showed that under the optimal conditions of other conditions, the K_m of enzyme was 0.67 mmol / L , K (cat) / Km was 13.53 mmol L ~ (-1) s ~ (-1) .Afterwards, 55 candidate AChE inhibitory compounds were screened from the SPECS chemical library and 9 antibacterial activities were obtained, The AChE inhibitory compounds of verotrophin from Tetranychus cinnabarinus and their toxicity to Tetranychus cinnabarinus are very good.This study provides a theoretical basis and experimental basis for the screening of acetylcholinesterase inhibitors and the development of pesticides targeting acetylcholinesterase basis.