Integrated bioprocess for the production and purification of recombinant proteins by affinity chromatography in Escherichia coli

  • Usama BeshayEmail author
  • Gerhard Miksch
  • Karl Friehs
  • Erwin Flaschel
Original Paper


In order to improve the effectiveness of the production of recombinant proteins in E. coli, integrated fermentation processes were developed. Therefore, expression vectors were constructed containing a strongly expressed gene for a β-glucanase fused with a metal-chelating affinity tag and a leader peptide for directing the fusion protein into the periplasmic space. Its export into the medium was achieved by means of co-expression of a bacteriocin-release protein, the Kil protein from pColE1. Bioreactors were modified so that special devices containing metal chelate pentadentate chelator PDC resins were located within the bioreactor. Using the bioreactor with an internal device the Zn2+-PDC had a 4.3-fold higher binding capacity than metal-free PDC (12.3 and 2.6 kU ml−1 PDC, respectively. Using the bioreactor with charged PDC in an external circuit revealed even higher β-glucanase concentration (65.6 kU ml−1), i.e. 1.5-fold compared to the internal adsorbent system.


Integrated process Production and purification E. coli β-glucanase 

List of symbols


mass concentration (g/l)


extracellular glucanase activity (U/ml)


adsorbed glucanase activity on resin (U/ml)


total glucanase activity (U/ml)


time of cultivation (h)



Dr. U. Beshay acknowledges the financial support from the Alexander von Humboldt Foundation for a visiting fellowship to Bielefeld University, Germany.


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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Usama Beshay
    • 1
    Email author
  • Gerhard Miksch
    • 2
  • Karl Friehs
    • 2
  • Erwin Flaschel
    • 2
  1. 1.Bioprocess Development DepartmentMubarak City for Scientific Research and Technology Applications, Genetic Engineering and Biotechnology Research Institute (GEBRI)AlexandriaEgypt
  2. 2.Technische FakultaetUniversitaet BielefeldBielefeldGermany

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