Purification of acetylcholinesterase from Halyomorpha halys (Stal) (Heteroptera: Pentatomidae) and management of control of the Pest through inhibition of the enzyme

Abstract

Halyomorpha halys (Stal) (Heteroptera: Pentatomidae) has brought about agricultural harm throughout the Eastern Black Sea coastline since 2017. It continues to come to a threat because there are no adequate studies on managing this pest. One of the major rein strategies of insecticides is acetylcholinesterase (ACHE) inhibition. Therefore, this study aims to investigate an alternative way to struggle H. halys by inhibiting ACHE. The ACHE was purified from H. halys using edrophonium-Sepharose 6B affinity chromatography and characterized by examining some kinetic properties. The molecular weight of the purified enzyme was determined by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and natural polyacrylamide gel electrophoresis. In addition, the inhibitory effects of tacrine and edrophonium chloride, and water extracts of olive leaf, walnut leaf and alder leaf on this ACHE were investigated. The acetylcholinesterase was purified 403-fold with an 83.3% yield. H. halys ACHE was found to have six subunits and a molecular weight of approximately 350 kDa. The ACHE's K-m, V-max, and k(cat) values were assigned to be 0.02 & PLUSMN; 0.006 mM, 3,333.3 & PLUSMN; 481 EU.mg protein(- 1), and 1070.2 & PLUSMN; 184 min(- 1), respectively. All inhibitors highly inhibited of activity of h.halys ACHE. Especially, the fact that the water extracts of these plants are effective in ACHE inhibition is significant due to being environmentally amicable pesticides that may be benefited in the struggle with the pest.