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  • Title: Overexpression of the phytase from Escherichia coli and its extracellular production in bioreactors.
    Author: Miksch G, Kleist S, Friehs K, Flaschel E.
    Journal: Appl Microbiol Biotechnol; 2002 Sep; 59(6):685-94. PubMed ID: 12226725.
    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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