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Research Journal of Microbiology
  Year: 2018 | Volume: 13 | Issue: 1 | Page No.: 47-52
DOI: 10.3923/jm.2018.47.52
 
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Extraction of Antigenic Membrane Proteins from Salmonella Using Detergent and Phase Partition Method
Venkatesh Padmanabhan, G. Arun Govind, A.S. Kamalanathan and N.S. Jayaprakash

Abstract:
Background and Objective: Salmonella porins have been shown to be good immunogens and these proteins carry an important value as they can be utilized as target proteins for diagnostics. In general, purification of porins from outer membrane complexes of bacteria is a tedious task due to the presence of enormous amounts of lipopolysaccharides and flagellar proteins associated with the outer membrane complex along with the porin proteins. The objective of study was to employ a modified method using a combination of detergent and phase separation to get a good efficacy of the classical procedure. Materials and Methods: The Salmonella enterica strain was obtained from MTCC, IMTECH-Chandigarh, India. The bacterial cultures were grown in standard nutrient broth and harvested for extraction of outer membrane proteins with Zwittergent™ 3-14 and phase separation using Triton X-114 (TX-114). The extracted protein was analyzed and confirmed using ESI-QTOF mass spectrometry. Results: Varying percentage of sodium chloride and glycerol were tried out and all the phase separated profiles gave the same result with varying protein concentrations of the outer membrane porin (Omps) and Lipopolysaccharide (LPS) in the upper and lower phase. A good protein profile in terms of absence of LPS and protein enrichment was obtained when 5% sodium chloride and 5% glycerol was used. The proteins identified by mass spectrometric analysis matched predominately to Salmonella species. These proteins gave unique peptide matches for outer membrane proteins (OMPs) aided by NCBI database. Conclusion: The modified detergent extraction protocol along with phase separation was effective in extracting the membrane proteins from Salmonella sp.
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How to cite this article:

Venkatesh Padmanabhan, G. Arun Govind, A.S. Kamalanathan and N.S. Jayaprakash, 2018. Extraction of Antigenic Membrane Proteins from Salmonella Using Detergent and Phase Partition Method. Research Journal of Microbiology, 13: 47-52.

DOI: 10.3923/jm.2018.47.52

URL: https://scialert.net/abstract/?doi=jm.2018.47.52

 
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