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Cloning of ethyl carbamate hydrolase (ECH) and its overexpression in E. coli. Lane 1, E. coli BL21 (DE3); lane 2, whole cells of E. coli BL21 (DE3)/pET24b-ECH induced with IPTG; lane 3, puncture fluid; lanes 4 to 6, ECH was not eluted from a crude enzyme solution with different concentration of imidazole (20, 100, 200 mM); lane 7, ECH was eluted from crude enzyme solution with 300 mM imidazole solution (33 kDa); lane M: protein marker.
