Overexpression of the phytase from Escherichia coli and its extracellular production in bioreactors
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- Miksch, G., Kleist, S., Friehs, K. et al. Appl Microbiol Biotechnol (2002) 59: 685. doi:10.1007/s00253-002-1071-z
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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.