Biosynthesis of the Escherichia coli siderophore enterobactin: sequence of the entF gene, expression and purification of EntF, and analysis of covalent phosphopantetheine

Biochemistry. 1991 Mar 19;30(11):2916-27. doi: 10.1021/bi00225a027.


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.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 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


  • DNA, Bacterial
  • Multienzyme Complexes
  • Enterobactin
  • Pantetheine
  • Amino Acid Isomerases
  • Peptide Synthases
  • gramicidin-S-synthetase 2
  • tyrocidine synthetase
  • 2,3-dihydroxybenzoate - serine ligase
  • 4'-phosphopantetheine

Associated data

  • GENBANK/J05325