PMID:12226725

Citation	Miksch, G, Kleist, S, Friehs, K and Flaschel, E (2002) Overexpression of the phytase from Escherichia coli and its extracellular production in bioreactors. Appl. Microbiol. Biotechnol. 59:685-94
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.
Links	PubMed Online version:10.1007/s00253-002-1071-z
Keywords	6-Phytase/biosynthesis; 6-Phytase/chemistry; 6-Phytase/genetics; 6-Phytase/metabolism; Acid Phosphatase/chemistry; Acid Phosphatase/genetics; Acid Phosphatase/metabolism; Amino Acid Sequence; Base Sequence; Bioreactors; DNA, Bacterial/chemistry; DNA, Bacterial/genetics; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/genetics; Fermentation; Kinetics; Molecular Sequence Data; Mutation; Polymerase Chain Reaction; Recombinant Proteins/chemistry; Recombinant Proteins/genetics
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