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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.
*Corresponding author. E-mail:
prasadjavarappa@hotmail.com.
Accepted
10
March, 2009 |
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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. |
