E. coli is one of the most widely used hosts for production of recombinant polypeptides, including therapeutic proteins. However, protein mis-folding and aggregation, proteolysis, and the lack of disulfide bond formation often make this process challenging in producing a functional protein.
Targeting recombinant protein production to the periplasmic space of Escherichia coli presents several advantages over cytoplasmic production in inclusion bodies and at the same time overcomes the low productivity problem often associated with culture medium secretion. Isolation of proteins from periplasm is easier than from whole cell lysates. Periplasm also provides an oxidizing environment where the disulfide bond formation (Dsb) system catalyzes the formation of disulfide bonds. Therefore, periplasm proves a favorable compartment for production of disulfide bond containing proteins like antibody fragments, and many peptide hormones, enabling their folding into native conformation. Read More
