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Gene Review

entF  -  enterobactin synthase subunit F

Escherichia coli O157:H7 str. Sakai

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Disease relevance of ECs0625


High impact information on ECs0625

  • We report the use of combinatorial mutagenesis coupled with in vivo selection for the production of the Escherichia coli NRPS product enterobactin to map the surface of the aryl carrier protein (ArCP) domain of EntB that interacts with the downstream elongation module EntF [3].
  • BACKGROUND: EntF is a 142 kDa four domain (condensation-adenylation-peptidyl carrier protein-thioesterase) nonribosomal peptide synthetase (NRPS) enzyme that assembles the Escherichia coli N-acyl-serine trilactone siderophore enterobactin from serine, dihydroxybenzoate (DHB) and ATP with three other enzymes (EntB, EntD and EntE) [4].
  • 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 [5].
  • SIGNIFICANCE AND IMPACT OF THE STUDY: The potential benefit of having an expression system based on EntF is that gene expression can be finely controlled upon addition of low concentrations of a peptide that can easily be artificially synthesized [6].

Associations of ECs0625 with chemical compounds

  • RESULTS: The Ser1138-->Cys mutant (kcat decreased 1000-fold compared with wild-type EntF) releases both enterobactin (75%) and linear (DHB-Ser)2 dimer (25%) as products [4].
  • Release of PPi during incubation of EntF with serine and ATP was observed, but with a low turnover number of 1.0 min-1 [7].


  1. The EntF and EntE adenylation domains of Escherichia coli enterobactin synthetase: sequestration and selectivity in acyl-AMP transfers to thiolation domain cosubstrates. Ehmann, D.E., Shaw-Reid, C.A., Losey, H.C., Walsh, C.T. Proc. Natl. Acad. Sci. U.S.A. (2000) [Pubmed]
  2. Vibrio cholerae VibF is required for vibriobactin synthesis and is a member of the family of nonribosomal peptide synthetases. Butterton, J.R., Choi, M.H., Watnick, P.I., Carroll, P.A., Calderwood, S.B. J. Bacteriol. (2000) [Pubmed]
  3. A protein interaction surface in nonribosomal peptide synthesis mapped by combinatorial mutagenesis and selection. Lai, J.R., Fischbach, M.A., Liu, D.R., Walsh, C.T. Proc. Natl. Acad. Sci. U.S.A. (2006) [Pubmed]
  4. Assembly line enzymology by multimodular nonribosomal peptide synthetases: the thioesterase domain of E. coli EntF catalyzes both elongation and cyclolactonization. Shaw-Reid, C.A., Kelleher, N.L., Losey, H.C., Gehring, A.M., Berg, C., Walsh, C.T. Chem. Biol. (1999) [Pubmed]
  5. Biosynthesis of the Escherichia coli siderophore enterobactin: sequence of the entF gene, expression and purification of EntF, and analysis of covalent phosphopantetheine. Rusnak, F., Sakaitani, M., Drueckhammer, D., Reichert, J., Walsh, C.T. Biochemistry (1991) [Pubmed]
  6. Potential of the enterocin regulatory system to control expression of heterologous genes in Enterococcus. Hickey, R.M., Twomey, D.P., Ross, R.P., Hill, C. J. Appl. Microbiol. (2003) [Pubmed]
  7. Characterization of EntF as a serine-activating enzyme. Reichert, J., Sakaitani, M., Walsh, C.T. Protein Sci. (1992) [Pubmed]
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