i am working on a project about periplasmic expression of a protein with secB-dependent pathway in origami DE3. Although i have recombinant protein in total cell fraction, periplasmic fraction doesn't contain my secretory protein! may origami as expression host interfere with secretion pathway? is it possible? i would be grateful if you could recommend any efficient bacterial expression system for secretion of my protein. thanks in advance.
SecB-dependant proteins are secreted while still unfolded (that is one of the roles of SecB). If the structure of your protein depends on the presence of correctly formed disulfide bridges, it might be already folded by the time it reaches the membrane. I would try another host; plain BL21(DE3) should do.
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