ISOLATION, PURIFICATION AND BIOPHYSICAL CHARACTERIZATION OF BASIC 7S GLOBULIN FROM COCOS NUCIFERA
Journal Title: Int J of Pharm Res & Analy - Year 2015, Vol 5, Issue 2
Abstract
Elucidation of structure-function relationships of seed storage proteins is a prerequisite for developing theoretically new food and/or food materials based on seed storage proteins. Furthermore, recent findings suggest that the 7S globulin, among all storage proteins in the coconut seeds, is responsible for the up-regulation of LDL receptors and that this activation is induced by the α and αβ- subunits of 7S globulin. The molecular mechanism underlying this biological response is currently under investigation. Here we report the purification and biophysical characterization of 7S globulin protein from the coconut endosperm. The total protein was separated by centrifugation at 13500g for 15 min at 4 ºC. The total precipitated protein of the 60% ammonium sulfate was subjected to column chromatography on a Hi-Prep 1.5/20 DEAE Sephadex A-50 HR column at a flow rate of 1 mL/min followed by purification using gel filtration chromatography. The purified fractions will be analyzed by SDS-PAGE. The band of SDS PAGE was excised and identified by mass spectrometry. These fragments are analyzed through MASCOT and identified through BLAST searches. We further perform circular dichroism (CD) analysis to observe the effect of temperature on secondary structure of the protein and thermal denaturation studies were carried out at 222 nm. This study will provide a sound basis for understanding in the structure-function relationship of coconut protein especially basic 7S globulin.
Authors and Affiliations
Alpana Kumari
Keywords
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