AJ Life Sci. 2021, 4 (1): 127-135

Isolation and Purification of Thrombolytic Enzyme Extracted from Earthworm Punjab, Pakistan

Bakhtawer *1,Muhammad Faheem malik1 and Sumera Afsheen 1

1 Department of zoology, University of Gujrat, Punjab, Pakistan



Abstract

The Cardiovascular disease due to thrombus (clot) formation is the major factor of death throughout the world. Earthworms being the eco engineers has thrombolytic enzyme that can be used for thrombolysis. The thrombolytic enzyme was isolated and purified from supernatant of earthworm Apporectodea longa by column chromatography. Six Strain BKT 11, BKT 15, BKT 17, BKT 26, BKT 27 and BKT 28 shows the thrombolytic activity 791.64 U/mg, 1362.39 U/mg, 1205.4 U/mg, 710.63 U/mg, 529.66 U/mg and 625.00 U/mg respectively. Thrombolytic activity was confirmed by blood clot lysis method. Different concentrations 50 ?l,100 ?l, 150 ?l, 200 ?l and 250 ?l of extracted enzyme were applied on 25mg of wet blood clot along with control where distill water used. These fractions of extracted enzymes represent the dissolution of clot (thrombolysis). The molecular weight 32 KDa was determined by sodium dodecyl sulphate gel electrophoresis (SDS-PAGE). Results show that extracted elute have potential of fibrinolytic activity in this specie of earthworm and it can serve as a suitable therapeutic agent.

Keywords: Thrombolytic activity, Casein plate assay, Blood clot lysis, spectrophotometry, Gel electrophoresis.

DOI

https://doi.org/10.34091/AJLS.4.1.15

Received

September 5, 2020

Received Revised

January 10, 2021

Accepted

June 26, 2021

Available Online

September 11, 2021


Corresponding author email:

Bakht.warraich99@gmail.com

How to Cite

Abasyn Journal of Life Sciences , ISSN (online): 2663-1040, Published by Abasyn University