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. Author manuscript; available in PMC: 2014 Feb 1.
Published in final edited form as: Mol Microbiol. 2012 Dec 19;87(3):594–608. doi: 10.1111/mmi.12118

Figure 2.

Figure 2

Cee is an enterobactin esterase located in periplasm of C. jejuni. (A) SDS-PAGE analysis of rCee production and purification. Lane 1, molecular weight marker (Bio-Rad); lane 2, whole cell lysate of noninduced E. coli; lane 3, whole-cell lysate of E. coli induced for 3h with 1 mM IPTG; lane 4, purified rCee purified. (B) Localization of Cee. The proteins samples were subjected to SDS-PAGE and immunoblotting analysis using CmeR as a cytoplasmic protein control and CmeB as an inner membrane control. Lane 1, whole cell lysate of C. jejuni 81–176 containing control vector pRY111 (JL229); lane 2, whole cell lysate of the C. jejuni 81–176 containing a vector that overproduces Cee (JL870); lane 3, periplasmic fraction of JL870; lane 4, spheroplastic fraction of JL870; and lane 5, the purified rCee. (C) Thin layer chromatography (TLC) analysis of the hydrolysis of Ent by rCee. The reaction time was indicated above the picture. Inline graphic, Ent, or cyclic trimer of DHBS; Inline graphic, linear trimer of DHBS; Inline graphic, dimer of DHBS; Inline graphic, DHBS monomer. (D) HPLC analysis of hydrolysis of apo Ent by Cee, Fes, IroD, and IroE. (E) HPLC analysis of hydrolysis of FeEnt by Cee, Fes, IroD, and IroE.