ISSN 2756-3650
International Journal of Animal Breeding and Genetics ISSN 2756-3650, Vol. 14 (2), pp. 001-008, February, 2025. Available online at www.internationalscholarsjournals.org © International Scholars Journals
Full Length Research Paper
Isolation and Characterization of Acetylcholinesterase Enzyme from Pardosa astrigera L. Koch
Min Ma1*, Bin Zhang2 and Sheng-cai Li1
1College of Agriculture, Shanxi Agricultural University, Taigu 030801, Shanxi, China.
2College of Arts and Science, Shanxi Agricultural University, Taigu 030801, Shanxi, China.
Accepted 10 January, 2025
A simple and effective method was set up to purify acetylcholinesterase (AChE, EC3.1.1.7) from the head and appendage of Pardosa astrigera. The AChE was purified by salt fractionation, DEAE -52 ion-exchange chromatography and Superdex 200 methods. The purity and molecular weight of the acetylcholinesterase were detected with sodium dodecyl sulphate-polyacrylamide gel (SDS-PAGE) electrophoresis. Biochemical properties were measured from the purified AChE by improves Ellmans method. The AChE was extracted from the head and appendage of Pardosa astrigera. SDS- PAGE electrophoresis showed a single band with relative molecular weight of 66.35 KDa. However, three bands resolved on PAGE gel electrophoresis, leading to the inference that native AChE exists in three forms. AChE purification fold can be reached to 229.60 when the yield was 13.88%. Significantly highest purified AChE activity was observed when the experiment conditions were 30oc, pH 7.5, 700 umol.L-1 ATChI. IC50, the half maximal inhibitory concentration, represents the concentration of an inhibitor that is required for 50% inhibition of things. The IC50 values ratios of crude extract to purified AChE were 6.54, 7.52, 3.92 and 5.08 times respectively, which was caused by methomyl, phoxim, beta cypermethrin and chlorpyrifos. We got the AchE of the electrophoretically purity from the head and appendage of Pardosa astrigera, and the sensitive of purified enzyme was restrained more easily.
Key words: Salt fractionation, chromatography, characterization.