Isolation and Purification of Acetylcolinesterases (AchE ) from Blood of Thalassimic Patient and Kinetic Studies for Purified Enzyme

Nazar Ahmed Naji, Sayran Sattar Saleh, Galawesh Noori Taher

Abstract


In this research to isolate and study the properties of the Acetylcolinesterase  (Ec 3 .1.1.7) enzyme in the blood of a patient suffering from β- thalassemia major and study kinetic  studies for purified  AchE .The study involves taking(5ml) of crude serum of thalassemic patients and subjected to a series of purification  processes including : precipitation by ammonium  sulfate , filtration by centrifugation radiator , dialaysis in presence of Tris-HCL,separation using the technology of gel and then Estimation approximate Molecular Weight of Partially Purified AchE using gel filtration technique and sodium dodecyl sulfate (SDS)-pag polyacrylamid gel electrophoresist.The result of the study indicates that AchE has an approximat molecular weight of 173720 Dalton.The kinetics of the enzyme were studied and the results showed that the maximum velocity was 7.8 µmol/min/mol and the Km was 0.1 M whil the optimum temperature of the enzyme was( 37  )and the optimum pH was( 7.4) .


Keywords


Acetylcolinesterase ; β- thalassemia ; Purification; Dialaysis.

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