South Asian Journal of Life Sciences
Muhammad Shahid Nadeem1*, Jalaluddin Azam Khan1, Bibi Nazia Murtaza2, Khushi Muhammad2, Abdul Rauf3
1Department of Biochemistry, King Abdulaziz University, Jeddah, Saudi Arabia; 2School of Biological Sciences, University of the Punjab, Lahore, Pakistan; 3Department of Zoology, University of Azad Jamu and Kashmir, Muzafarabad, Pakistan.
*Correspondence | Muhammad Shahid Nadeem, King Abdulaziz University, Saudi Arabia; Email: shahid.nadeem.hu@gmail.com
Abstract
Catalase is ubiquitous enzyme which catalyses the breakdown of hydrogen peroxide (H2O2). The aim of present study was to purify and characterize the liver catalase from Bubalus bubalis, which is an important domestic animal in South Asia. The enzyme was purified to electrophoretic homogeneity by selective acetone precipitations and anion-exchange chromatography based on DEAE-Sephadex Colum. Purified enzyme displayed specific activity of 35753 units per mg of protein with 30.5 % recovery and 123 fold purification. Optimum activity was measured at pH 7 and 30°C. The KM value of purified catalase was calculated as 58mM for H2O2. Gel-filtration chromatography experiments indicate that the purified enzyme was a tetramer with MW of about 240 kDa. There was no effect of 10 to 50mM EDTA on the activity of enzyme after incubation for 30 min at 4°C. Our study reveals the first report on the purification and properties of catalase from river buffalo which is an unexplored species for its proteins. The isolated enzyme with high yield and specific activity, offers an attractive alternate for the bovine and porcine enzymes used in clinical laboratories.
Keywords | Catalase, River buffalo, Ion-exchange chromatography
Editor | Muhammad Nauman Zahid, Quality Operations Laboratory, University of Veterinary and Animal Sciences, Lahore, Pakistan.
Received | July 08, 2015; Revised | October 21, 2015; Accepted | October 23, 2015; Published | December 23, 2015
Citation | Nadeem MS, Khan JA, Murtaza BN, Muhammad K, Rauf A (2015). Purification and properties of liver catalase from water buffalo (Bubalus bubalis). S. Asian J. Life Sci. 3(2): 51-55.
ISSN | 2311–0589
Copyright © 2015 Nadeem et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.