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


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) .


Acetylcolinesterase ; β- thalassemia ; Purification; Dialaysis.

Full Text:



– Weatherall, D.J. ( 2010) . "The inherited diseases of hemoglobin are an emerging global health burden" J.; Blood, 115 .pp: 4331,36.

- S. Sultan, S. M. Irfan, R. Zeeshan, J. Kaker, and A. Kidwai.(2014). “Zinc status and its correlation with basic parameters in transfusion dependent thalassemic patients: a Pakistani perspective,” Iranian Journal of Blood and Cancer, vol. 6, no. 3, pp. 113–118.

- Nasiri, m.; Hosseini,S.H.;and shah Mohammadi,S.(2014).Mental health status in patiemts with Thalassemia major in Iran,Journal of Pediatrics Review office ,2 (1):55-56 .

- Gupta, V.K, Pal,NJ. Siddiqi andB.Sharma,(2015).Acetylcholineserase From human erythrocytes as a surrogate biomarker of lead induced neurotoxicity .Enzyme Res, 10.1155/2015/370705.

- Jean P.Dzoyem ,Victor Kuete ,Jacobus N.(2014).Toxicological Survey of African Medicinal plants , p 659-715.

- Gupta, V.K, Pal,NJ. Siddiqi andB.Sharma,(2016).Carbofuran modulating functions of acetylcholinesterase from rat brain in vitro Adv. Biol,10.1155/2016/3760967.

- Zhang ,X.J., Greenbreg,D. S.(2012). Acetyl cholinesterase involvement in apoptosis. Front Mol Neuro Sci.,5:40.

- Moscoso,F.,Deive,FJ.,Esperanca,JS.and Rodriguez,A.2013.Pesticide Removal from Aqueous Solution by Adding Salting Out Agents Int.J.Mol,Sci.,14:20954- 20965 .

- Plummer,T.D. (1978). "An Introdaction of Practical Biochemistry". 2nd ed .; Mcgraw – Hill Book. ; Co . V. K.pp: 48, 52, 174, 270, 274.

- Garrett, Reginald H.; Grisham, Charles M. (2013). Biochemistry (5th ed.). Belmont, CA: Brooks/Cole, Cengage Learning. p. 108. ISBN 9781133106296.

- Burgess, R . R .and Dustscher,M . P.(2013).Guid to protein(purification and characterization),Method in Enzymology.,(463):320 .

- Berg, J. M., Tymoczko, J. L., Stryer, L. (2012). Biochemistry. 17th ed. W. H. Freeman and Company. New York. USA. pp. 68,69,138, 139, 145, 146.

- A.DrabikA.Bodzoń-KułakowskaJ.Silberring Proteomic Profiling and Analytical Chemistry (Second Edition) 2016, Pages 115-143.

- Ellman G. L.. Courtney K. p., Andres V. and Feather stone R. M., Bioche , Pharmacol, 1961, 7, 88-91

- Lowry, O. H., Rosbrough, N.J.,Farr, A. L.and Randall, R.J.1951. Protein measurement with the afolin phenol reagent , J Biol. Chem.;193:265.

- Murrqy,RK.,Bender,DA.,Botham,KM.,Kennelly,PJ.,Rodell,VW.,Weil, PA.2012.Harber Illustrated Biochemistry.30th ed.,The Mcgraw-Hill Companies,China.Chapter 8.p.209.

- Xi, H. J., Wu, R. P., Liu, J. J., Zhang, L. J., Li, Z. S.Role of acetyl cholinesterase in lung cancer. Thorac Cancer, 2015; 6(4): 390-398.

- Tripathi, A., Srivastava ,U.C. (2008). Acetylcholinesterase : Aversatile enzyme of enzyme of nervous system .Annals of Neurosclences ,Vol. No.4.

- Kaya, H. B. Özcan, B. Şişecioğlu, M. Ozdemir, H.(2013). Purification of acetylcholinesterase by 9-Amino-1,2,3,4-tetrahydroacridine from human erythrocytes .Applied Biochemistry and Biotechnology.170(1):198–209.

- Lelas F. B.(2017). Studying of SomeBiochemical Parameters in

Cerebrospinal Fluid for Children with Acute Lymphoblastic Leukemia and Hydrocephalus Ph. D. Thesis Submitted to College of Science. University of Mosul.

- McIntosh, C. S. H. & Plummer, D. T. (1973) Biochem. J. 133, 655-665.

- Levinson S. R. and Ellory J. C.(1974). The Molecular Form of Acetylcholinesterase as Determined by Iradiation Inactivation Biochem. J. 137, 123-125.

- Firas T.M .(2017).Biochemical study of novel phosphate paracetamol prodrug invivo and invitro. Journal of university of Anbar for pure Science.,vol 11(2) ,p 12-29 .

- Vivek Kumar Gupta, Abhishek Kumar1, Nikhat Jamal Siddiqi and Bechan Sharma.(2016)." Rat Brain Acetyl Cholinesterase as a Biomarker of Cadmium Inducecity" Journal of Toxicology Volume (1) Issue (1).

- Vivek Kumar Gupta,Abhishek Kumar, Sarita H. Yadav, Rashmi Pandey and Bechan Sharma.(2017)." Acetylcholinesterase as a Biomarker of Arsenic Induced Cardiotoxicity in Mammals"DOI: 10.17311/sciintl..142.149.

- Ahmed M , Latif N , Khan RA , Ahmad A , Rocha JBT , Mazzanti CM , Bagatini MD ,Morsch VM , Schetinger MRC .(2012). Enzymatic and biochemical characterization of Bungarus sindanus snake venom acetylcholinesterase.J Venomous Animals and Toxins including Tropical Diseases ., volume 18 | issue 2 | pages 236-243.


  • There are currently no refbacks.





About IJSBAR | Privacy PolicyTerms & Conditions | Contact Us | DisclaimerFAQs 

IJSBAR is published by (GSSRR).