Applied Microbiology and Biotechnology

, Volume 59, Issue 6, pp 685–694

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

Original Paper

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.

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Copyright information

© Springer-Verlag  2003

Authors and Affiliations

  1. 1.Department of Fermentation Engineering, Faculty of Technology, University of Bielefeld, 33501 Bielefeld, GermanyGermany

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