Expression condition optimization and product structure analysis of human p-defensin-2 fusion protein in engineered bacteria

摘要

Using conical flask culture for simulation of small-scale fermentation, we studied the changes in β-defensin-2 (HBD-2) protein expression when HBD-2-harbording engineered bacteria were exposed to different culture conditions, culture media, incubation temperatures, pH values, and inducer concentrations. We also performed computer simulation-based structural analysis of the target protein. Our results showed that HBD-2 fusion protein was most abundantly expressed in the engineered bacteria cultured in M_9B_2 medium at 32°C at a pH range of 7.3 to 7.5 upon induction by 1.2 mmol/L IPTG. Computer simulation-based structure analysis showed that enhanced hydrophilicity of the recombinant protein did not reduce the epitopes of HBD-2 fusion protein and that HBD-2 fusion protein kept the same structure as that of the native protein.