Development of extracellular production system of recombinant proteins in recombinant Escherichia coli

Background

Escherichia coli has been the workhorse for the production of various recombinant proteins and metabolites because of the availability of well established technologies for genetic manipulation and cultivation. Various strategies have been employed for the development of E. coli strains which are able to efficiently produce recombinant proteins [1, 2]. Extracellular production of recombinant proteins has advantages over secretion into the periplasm [2]. Extracellular production does not require outer membrane disruption to recover target proteins, and therefore, it avoids intracellular proteolysis by periplasmic proteases and allows continuous production of recombinant proteins, and new approaches for the extracellular production of recombinant proteins in E coli are discussed.

Results

E. coli BL21 strains showed the high accumulation of OmpF protein in culture medium during high cell density cultivation (see Figure 1). From this interesting phenomenon, a new and efficient method for the extracellular production of recombinant protein in E coli was developed. Using this new developed extracellular production system, various recombinant proteins could be efficiently produced into culture medium.

Figure 1

SDS-PAGE analysis of culture supernatant from the high cell density culture of E. coli BL21(DE3).

Conclusion

We developed new approaches for the extracellular production of recombinant proteins in E. coli by using one of the outer membrane proteins as a partner.