Constitutive expression of protective antigen gene of Bacillus anthracis in Escherichia coli

Biochemical and Biophysical Research Communications
V ChauhanR Bhatnagar

Abstract

The fatal bacterial infection caused by inhalation of the Bacillus anthracis spores results from the synthesis of protein toxins-protective antigen (PA), lethal factor (LF), and edema factor (EF)--by the bacterium. PA is the target-cell binding protein and is common to the two effector molecules, LF and EF, which exert their toxic effects once they are translocated to the cytosol by PA. PA is the major component of vaccines against anthrax since it confers protective immunity. The large-scale production of recombinant protein-based anthrax vaccines requires overexpression of the PA protein. We have constitutively expressed the protective antigen protein in E. coli DH5alpha strain. We have found no increase in degradation of PA when the protein is constitutively expressed and no plasmid instability was observed inside the expressing cells. We have also scaled up the expression by bioprocess optimization using batch culture technique in a fermentor. The protein was purified using metal-chelate affinity chromatography. Approximately 125 mg of recombinant protective antigen (rPA) protein was obtained per liter of batch culture. It was found to be biologically and functionally fully active in comparison to PA protein from Bacillus a...Continue Reading

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Citations

Jul 26, 2008·Applied Biochemistry and Biotechnology·Fatemeh VahediMahmoud Mahmoudi
Oct 24, 2007·Journal of Immune Based Therapies and Vaccines·Cassandra D KellyNick M Cirino
Mar 17, 2004·Biochemical and Biophysical Research Communications·Samer SinghRakesh Bhatnagar
Sep 9, 2011·Journal of Industrial Microbiology & Biotechnology·Hendrik Waegeman, Wim Soetaert
Feb 25, 2003·Protein Expression and Purification·Sangeeth KrishnanchettiarMichael Caffrey
Mar 12, 2016·Molecular Biology International·Nagendra SuryanarayanaUrmil Tuteja
Sep 1, 2004·Biochemical and Biophysical Research Communications·Samer SinghRakesh Bhatnagar
Oct 13, 2004·Protein Expression and Purification·Michael W LairdWilliam Gwinn
Jul 6, 2015·International Journal of Medical Microbiology : IJMM·Somya AggarwalRakesh Bhatnagar
Apr 10, 2007·Biochemical and Biophysical Research Communications·Subhash ChandraRakesh Bhatnagar
Jun 27, 2006·Journal of Biotechnology·Hsin-Hou ChangDer-Shan Sun
Nov 26, 2002·Biochemical and Biophysical Research Communications·Mohd Azhar AzizRakesh Bhatnagar
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Jul 13, 2019·Expert Review of Vaccines·Olga A KondakovaOlga V Karpova
Nov 19, 2005·The Journal of Biological Chemistry·Roman A MelnykR John Collier
Sep 5, 2002·Biochemical and Biophysical Research Communications·Samer SinghRakesh Bhatnagar

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