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Purification of toxin protein from E. coli isolate pe88

Author Affiliations

  • 1Dept. of Microbiology, Centre for P.G. Studies, 18/3, 9th Cross, 3rd Block, Jayanagar, Jain University, Bangalore-11, India
  • 2Dept. of Microbiology, Centre for P.G. Studies, 18/3, 9th Cross, 3rd Block, Jayanagar, Jain University, Bangalore-11, India

Int. Res. J. Biological Sci., Volume 5, Issue (6), Pages 57-59, June,10 (2016)

Abstract

Escherichia coli isolate pe88, which showed positive to Stx2c toxin by PCR analysis was selected for the present study. The protein from E. coli culture was subjected to salting out by ammonium sulphate precipitation till the saturation followed by dialysis using 1% sucrose solution for overnight. Further purification of protein from the isolate was carried out by ion-exchange chromatography. Column was developed using DEAE Cellulose for anion and Sephrose for cation exchanger. Elution buffer was used as mobile phase. Procedures were performed at room temperature. Elutions collected every five minutes were subjected to protein estimation by Lowry’s method using Bovine Serum Albumin as standard. The elution of protein from the isolate pe88 was subjected to SDS-PAGE using standarded molecular weight marker. The sample showed a single band close to 74 kDa after purification by column chromatography.

References

  1. Hodson A.W. and Latner A. (1961)., Isoenzymes of Alkaline Phosphate., British Medical Journal., 2(5246), 243.
  2. Faiza A., Ali and Nada Hamza B. (2012)., Purification and characterization of Alkaline Phosphate enzyme from the periplasmic space of Escherichia coli C90 using different methods., Africian Crop Science Journal, 20(2), 125-135.
  3. David L., Macleod and Carlton L. Gyles (1990)., Purification and characterization of Escherichia coli Shiga-like toxin II variant., Americian society for Microbiology, 58(5), 1232-1239.
  4. Lowry O.H., Rosebrought N.J., Farr A.L and Randall R. J. (1951)., Protein measurement with the Folin phenol reagent., J. Biol. Chem., 193, 265 (The orginal).
  5. Sadasivam S. and Manikam A. (1990)., Biochemical methods., Third edition vol 2, p 54-59 New age international limited publishers, New Delhi.
  6. Brien ADO and Gerald D. Laveck (1983)., Purification and characterization of Shigella dysenteriae 1- like toxin produced by Escherichia coli., American society for Microbiology, 40(2), 675-683.