Inhibition of acetylcholinesterase (AChE) is one of the important modes of action available for the control of insects. An assessment was made of the electric eel AChE inhibitory activity of 55 endophytic fungi isolated from the leaves of Huperzia serrata collected in Eunshi, Hubei Province, China. Of 55 mycelial extracts and 55 ethyl acetate-soluble fractions from liquid culture of the fungi, the ethyl acetate-soluble fraction of strain ES056 exhibited strong AChE inhibitory activity (88%) at a concentration of 100 μg/mL. Based on morphological characteristics and nuclear ribosomal DNA ITS sequence analysis, this fungus showed greatest similarity to members of the order Eurotiales and was shown to be most closely related to members of the Aspergillus aculeatus group. It was designated Aspergillus sp. MY056. This strain merits further study as a potential anti-AChE product.