IJCRR - 5(16), August, 2013
A METHOD FOR EFFICIENT EXTRACTION OF CELL WALL PROTEINS FROM CANDIDA ALBICANS
Author: Pavithra Amrath Jain, R. D. Kulkarni, B.M. Swamy, Gopala Mugeraya, Srinikethan G., Sharanbasu K.A.
Category: General Sciences
Candida cell wall proteomics is a challenging area of research. Extraction of cell wall proteins needs to be done carefully to obtain reproducible results. We used two different methods for extraction of candida cell wall proteins and documented the modifications required. In the course of extraction, we also evaluated efficacy of cell wall disruption by mechanical procedure of homogenization with glass beads to increase the protein yield and purity. For extraction of candida cell wall proteins, we utilized detergents like Sodium dodecyl sulphate (SDS), dithiothreitol (DTT), tris base and chitinase. This method was further optimized using homogenization of candida cells with glass beads over different time intervals. Amount of cell disruption at each time interval of bead beating using two varieties of beads was evaluated by gram stain, culture, protein estimation and SDS PAGE subsequently. We observed that method using detergents and Chitinase yielded better quality proteins, less contaminated with salts. For homogenization, bead beating with glass beads of 0.5 mm diameter over 18 minutes was found to be superior. Our initial experiments revealed limits of both the methods in candida cell wall protein extraction. Mechanical disruption of the cells could be further optimized using routinely available vortex mixer to increase final protein yield and gave consistent and reproducible results.
Keywords: Candida albicans, Cell wall proteins, glass bead homogenization, SDS PAGE.
Pavithra Amrath Jain, R. D. Kulkarni, B.M. Swamy, Gopala Mugeraya, Srinikethan G., Sharanbasu K.A.. A METHOD FOR EFFICIENT EXTRACTION OF CELL WALL PROTEINS FROM CANDIDA ALBICANS International Journal of Current Research and Review. 5(16), August, 01-07
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