PMID:1826089
| Citation |
Rusnak, F, Sakaitani, M, Drueckhammer, D, Reichert, J and Walsh, CT (1991) Biosynthesis of the Escherichia coli siderophore enterobactin: sequence of the entF gene, expression and purification of EntF, and analysis of covalent phosphopantetheine. Biochemistry 30:2916-27 |
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| Abstract |
The sequence of the entF gene which codes for the serine activating enzyme in enterobactin biosynthesis is reported. The gene encodes a protein with a calculated molecular weight of 142,006 and shares homologies with the small subunits of gramicidin S synthetase and tyrocidine synthetase. We have subcloned and overexpressed entF in a multicopy plasmid and attempted to demonstrate L-serine-dependent ATP-[32P]PPi exchange activity and its participation in enterobactin biosynthesis, but the overexpressed enzyme appears to be essentially inactive in crude extract. A partial purification of active EntF from wild-type Escherichia coli, however, has confirmed the expected activities of EntF. In a search for possible causes for the low level of activity of the overexpressed enzyme, we have discovered that EntF contains a covalently bound phosphopantetheine cofactor. |
| Links | |
| Keywords |
Amino Acid Isomerases/genetics; Amino Acid Sequence; Base Sequence; Chromatography, Gel; Chromatography, Ion Exchange; Cloning, Molecular; DNA, Bacterial/genetics; DNA, Bacterial/isolation & purification; Enterobactin/metabolism; Escherichia coli/genetics; Escherichia coli/metabolism; Genes, Bacterial; Molecular Sequence Data; Molecular Weight; Multienzyme Complexes/genetics; Pantetheine/analogs & derivatives; Pantetheine/analysis; Peptide Synthases/genetics; Peptide Synthases/isolation & purification; Plasmids; Restriction Mapping; Sequence Homology, Nucleic Acid |
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