Applied Microbiology and Biotechnology

, Volume 59, Issue 6, pp 685–694

Overexpression of the phytase from Escherichia coli and its extracellular production in bioreactors

Authors

    • Department of Fermentation Engineering, Faculty of Technology, University of Bielefeld, 33501 Bielefeld, Germany
  • S. Kleist
    • Department of Fermentation Engineering, Faculty of Technology, University of Bielefeld, 33501 Bielefeld, Germany
  • K. Friehs
    • Department of Fermentation Engineering, Faculty of Technology, University of Bielefeld, 33501 Bielefeld, Germany
  • E. Flaschel
    • Department of Fermentation Engineering, Faculty of Technology, University of Bielefeld, 33501 Bielefeld, Germany
Original Paper

DOI: 10.1007/s00253-002-1071-z

Cite this article as:
Miksch, G., Kleist, S., Friehs, K. et al. Appl Microbiol Biotechnol (2002) 59: 685. doi:10.1007/s00253-002-1071-z

Abstract.

The gene for phytase from Escherichia coli was sequenced and compared with the appA gene. It was found to be a mutant derivative of the appA gene. After fusion with a C-terminal His-tag, phytase was purified by affinity chromatography and the enzymatic properties were analyzed. To develop a system for overexpression and extracellular production of phytase in E. coli, factors affecting the expression and secretion such as promoter type, host strain and selection pressure were analyzed. Using a secretion system based on the controlled expression of the kil gene, the expression of phytase was improved and the enzyme was released into the culture medium at a high level. An effective fermentation strategy based on fed-batch operation was developed.

Copyright information

© Springer-Verlag  2003