Chandrasekhar et al., IJPSR, 2014; Vol. 5(7): 2763-2771. E-ISSN: 0975-8232; P-ISSN: 2320-5148 International Journal of Pharmaceutical Sciences and Research 2763 IJPSR (2014), Vol. 5, Issue 7 (Research Article) Received on 20 January, 2014; received in revised form, 06 March, 2014; accepted, 05 April, 2014; published 01 July, 2014 IDENTIFICATION OF E.COLI NISSLE 1917 PROTEINS BY USING 2-D GEL ELECTROPHORESIS UNDER THE INFLUENCE OF COCOS NUCIFERA SAP AND WINE K. Chandrasekhar, S. Sreevani and J. Pramoda Kumari* Dept. of Microbiology, S.V.University, Tirupati, Andhra Pradesh, India INTRODUCTION: Cocos nucifera sap is commonly called as cocoti sap. Sap is a juice collected from the cocoti palm plants. After fermentation, cocoti sap is converted into cocoti wine. It is a sweetish, milky white, effervescent alcoholic beverage with evolved CO 2 bubbles as mildly alcoholic beverage similar to beer. This alcohol gives sour smell and sulphur like odor may also be present. It acts as toxicants. Many scientists have analyzed the chemical and microbial properties of cocoti sap 1, 2, 3 . Escherichia coli Nissle 1917 is one of the probiotic present in gut flora of our body and served as a model organism for countless biochemical, biological, and biotechnological studies, since the completion of the E. coli genome-sequencing project. QUICK RESPONSE CODE DOI: 10.13040/IJPSR.0975-8232.5(7).2763-71 Article can be accessed online on: www.ijpsr.com DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.5(7).2763-71 E. coli Nissle 1917 has demonstrated to reduce intestinal inflammatory bowel disease (IBD) 4, 5 . Proteomics is regarded as a powerful approach as far as biochemical research is concerned because it directly studies the key functional components of biochemical systems, namely proteins 6 . Proteomics provides a powerful tool for analysing the various molecular mechanisms employed by plants, animals, insects, and microorganisms. In most cases protein samples are compared in healthy versus infected (or) treated 7 .Proteomics that aims at the determination of proteins constituents and their isoforms in a given sample 8 . 2-D clean-up kit facilitates the preparation of low conductivity samples suitable for isoelectric focusing (IEF) and 2-D gel electrophoresis. Additionally, the kit concentrates proteins from samples that are too dilute, allowing for higher protein loads that can improve spot detection. Keywords: E. coli Nissle, protein, 2-D electrophoresis, up and down regulation. Correspondence to Author: J. Pramoda Kumari Dept. of Microbiology, Sri Venkateswara University, Tirupati, Andhra Pradesh, India- 517502. E-mail: pramodakumarij@gmail.com ABSTRACT: Proteomics is the large scale study of proteins. Before going to analyse the differentially expressed proteins, through 2-D gel electrophoresis the proteins were extracted in a sample under stress. In the present study, we were isolated the protein samples of E. coli Nissle 1917 treated with cocoti sap and wine stress. The isolated proteins washed with 2-D clean up kit because it was very sensitive to salts and detergents. Quantify the protein concentration for loading the samples in 2-D electrophoresis. Proteins were separated by based on IEF and Molecular weight. By using Image master 2-D platinum 6.0 software programmes analyze the protein spots in the gel. We noticed over all 800 proteins in our gel, 370 protein spots were visualized clearly, in that considers 15 spots were isolated based on the regulation. Ten spots shows up regulation and remaining shows down regulation. One newly expressed protein was isolated from wine treated gel when compared to the control and also sap treated gels. Independent t-test was performed to analyze the significant difference between up and down regulation values. Further, this study helps to undergo the proteins identification and its functions through MALDI-TOF analysis.