Expression of the cytoplasmic domain of NodC as an active form in Drosophila S2 cells

NodC, a membrane protein that catalyzes the synthesis of the chitin oligosaccharide chain, was successfully produced in a soluble form. The truncated NodC gene encoding only the cytoplasmic domain that deletes the hydrophobic N-terminus expressed both cytoplasmic and secreted proteins in Drosophila Schneider 2 cells. The expressed protein maintained the ability to synthesize chitin oligosaccharides, primarily (GlcNAc)4, similar to the native membrane-bound NodC. This evidence suggests that only the large hydrophilic loop of NodC is efficient for enzymatic activity. Moreover, immobilizing the soluble NodC to a solid phase has no effect on the enzymatic activity. This, anchoring NodC is not necessary for its activity.