PMID:23278903

Citation	Zeng, X, Mo, Y, Xu, F and Lin, J (2013) Identification and characterization of a periplasmic trilactone esterase, Cee, revealed unique features of ferric enterobactin acquisition in Campylobacter. Mol. Microbiol. 87:594-608
Abstract	Ferric enterobactin (FeEnt) acquisition is a highly efficient and conserved iron scavenging system in Gram-negative bacteria. Recently, we have characterized two FeEnt receptors (CfrA and CfrB) in Campylobacter jejuni and C. coli, the enteric human pathogens that do not produce any siderophores. In this study, whole-genome sequencing and comparative genomic analysis identified a unique Ent trilactone esterase Cee (Cj1376) in C. jejuni. Genomic analysis and biochemical assay strongly suggested that Cee is the sole trilactone esterase in C. jejuni. Thin-layer chromatography and HPLC analyses showed high efficiency of the purified Cee to hydrolyse Ent. Three Cee homologues previously characterized from other bacteria (IroE, IroD and Fes) were also purified and analysed together with Cee, indicating that Cee, Fes and IroD displayed similar hydrolysis dynamics for both apo and ferric forms of Ent while IroE catalysed Ent inefficiently. Unlike cytoplasmic Fes and IroD, Cee is localized in the periplasm as demonstrated by immunoblotting using Cee-specific antibodies. Genetic manipulation of diverse Campylobacter strains demonstrated that Cee is not only essential for CfrB-dependent FeEnt acquisition but also involved in CfrA-dependent pathway. Together, this study identified and characterized a novel periplasmic trilactone esterase and suggested a new model of FeEnt acquisition in Campylobacter.
Links	PubMed PMC3556234 Online version:10.1111/mmi.12118
Keywords	Campylobacter jejuni/enzymology; Campylobacter jejuni/genetics; Campylobacter jejuni/metabolism; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Enterobactin/metabolism; Esterases/genetics; Esterases/isolation & purification; Esterases/metabolism; Genome, Bacterial; Hydrolysis; Kinetics; Lactones/metabolism; Models, Biological; Molecular Sequence Data; Periplasmic Proteins/genetics; Periplasmic Proteins/isolation & purification; Periplasmic Proteins/metabolism; Sequence Analysis, DNA
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
CAMCO:I2E6K4		GO:0016788: hydrolase activity, acting on ester bonds	ECO:0000315:		F		Fig. 1	complete CACAO 9095
CAMCO:I2E6K4		GO:0034613: cellular protein localization	ECO:0000314:		P		Figure 2	complete CACAO 9096
CAMCO:I2E6K4		GO:0016788: hydrolase activity, acting on ester bonds	ECO:0000314:		F		Figure 2	complete CACAO 9097
CAMCO:I2E6K4		GO:0010039: response to iron ion	ECO:0000315:		P		Figure 3	complete CACAO 9098

References

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