Knocking out Lpp gene of E.coli CLM37 strain and extracellular production of N -glycosylated recombinant proteins / 上海交通大学学报（医学版）

ABSTRACT

Objective • To construct the E.coli CLM37 strain with Lpp gene deletion and to study the production of N-glycosylated recombinant proteins in this E.coli strain. Methods • Firstly, Red homologous recombination system was used to knock out the Lpp gene from the genome of E.coli CLM37. And then, the growth curve was detected to study the effects of deleted Lpp gene on the growth states of E.coli strain. Finally, the vector pIG6-rFn3-Gly which expresses receptor protein and the vector pACYCpgl, which carries N-glycosylation gene cluster that derives from Campylobacter jejuni, were co-transformed into E.coli CLM37ΔLpp to investigate the extracellular production of N-glycosylated recombinant proteins. Results • The E. coli CLM37ΔLpp with Lpp gene deletion was obtained, and the extracellular production of N-glycosylated rFn3-Gly was successfully achieved in this strain. Compared with E. coli CLM37, the total amount of rFn3-Gly produced via extracellular production of E. coli CLM37ΔLpp increased about 4 times, and the glycosylation efficiency increased about 6 times. Conclusion • N-glycosylated rFn3-Gly was successfully produced via extracellular production in E. coli CLM37ΔLpp, and the production of interest glycoprotein and the glycosylation efficiency were improved.