ISSN 2736-1756
Advanced Journal of Microbiology Research Vol. 2017
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2017 International Scholars Journals
Full Length Research Paper
Purification of human serum paraoxonase: A simple and rapid method
A. J. Mahadesh Prasad*, K. Kemparaju, Elizabeth A. Frank and Cletus J. M. D’Souza
Department of Biochemistry Manasagangotri, University of Mysore, Mysore-570006, India.
Accepted 10 March, 2017
Abstract
Paraoxonase/arylesterases (EC.3.1.8.2) is an enzyme found tightly associated with high density lipoprotein particle in serum. Because of its unique enzyme activity, antioxidant property and its role as an anti atherosclerotic molecule, various methods are used for its purification from human serum. Methods involved in its purification are elaborate and complicated. Also the yield and final activity are highly variable. Here, we report a 2 step method of purification involving affinity chromatography on cibacron blue sepharose followed by gel filtration on sephadex G50.The final preparation was 27.7 fold purified compared with the serum and gave a single band in SDS-PAGE by silver staining.
Key words: Cibacron blue sepharose, paraoxonase, phenyl acetate, SDS-PAGE, silver staining.