Erythrocyte acetylcholinesterase (AChE) is a preferred biomarker for the detection of organophosphorus poisoning. Acetylthiocholine (ATCh) is the most popular substrate for the detection of AChE activity. However, oximolysis is a prominent feature with ATCh. In this context, we have searched alternative substrates for AChE using in silico tools for screening of a better substrate. The in silico approach was performed to understand the fitness and the Total Interaction Energy (TIE) of substrates for AChE. The alternative substrates for AChE were screened in terms of high Goldscore and favorable TIE in comparison to acetylcholine (ACh)-AChE complex and other relevant esterases. Among the screened substrates, 1-Naphthyl acetate (1-NA) exhibited the most favorable interaction with AChE in terms of highest TIE and corresponding high Goldscore. The Molecular Dynamic (MD) simulation of the 1-NA-AChE complex showed a stable complex formation over a period of 5 ns. The results obtained in the in silico studies were validated in vitro using pure erythrocyte AChE and hemolysate. We observed 1-NA to be a better alternative substrate for AChE than ATCh in terms of lower Km value. Its specificity appeared at least similar to ATCh. Therefore, we propose that 1-NA can be an attractive chromogenic substrate for the measurement of AChE activity, and it possess the potential to detect organophosphorus pesticide (OP) poisoning.

红细胞乙酰胆碱酯酶（AChE）是用于检测有机磷中毒的优选生物标志物。乙酰胆碱（ATCh）是检测AChE活性的最流行底物。但是，肟化是ATCh的突出特征。在这种情况下，我们已经使用计算机工具搜索了用于AChE的替代底物，以筛选更好的底物。将在硅片为了了解AChE底物的适应性和总相互作用能（TIE），采用了A.E.A.的方法。与乙酰胆碱（ACh）-AChE复合物和其他相关酯酶相比，根据高Goldscore和有利的TIE筛选了AChE的替代底物。在筛选的底物中，就最高TIE和相应的高Goldscore而言，乙酸1-萘酯（1-NA）与AChE表现出最有利的相互作用。1-NA-AChE配合物的分子动力学（MD）模拟显示了在5 ns的时间内稳定的配合物形成。在计算机研究中获得的结果已在体外验证使用纯红细胞AChE和溶血产物。我们观察到1-NA在较低的Km值方面是比AChE更好的AChE替代底物。其特异性似乎至少类似于ATCh。因此，我们建议1-NA可以作为一种有吸引力的生色底物，用于测量AChE活性，并且它具有检测有机磷农药（OP）中毒的潜力。