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.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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6-Phytase / biosynthesis*
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6-Phytase / chemistry
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6-Phytase / genetics
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6-Phytase / metabolism
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Acid Phosphatase / chemistry
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Acid Phosphatase / genetics
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Acid Phosphatase / metabolism
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Amino Acid Sequence
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Base Sequence
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Bioreactors*
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DNA, Bacterial / chemistry
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DNA, Bacterial / genetics
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Escherichia coli / enzymology*
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Escherichia coli / genetics
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Escherichia coli Proteins / chemistry
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Escherichia coli Proteins / genetics
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Fermentation
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Kinetics
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Molecular Sequence Data
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Mutation
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Polymerase Chain Reaction
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
Substances
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DNA, Bacterial
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Escherichia coli Proteins
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Recombinant Proteins
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Acid Phosphatase
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6-Phytase
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appA protein, E coli