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Improving the production of human‐like collagen by pulse‐feeding glucose during the fed‐batch culture of recombinant Escherichia coli

To increase the target protein production and reduce acetic acid accumulation during fed‐batch cultivation of recombinant Escherichia coli BL21 in a 30‐L bioreactor, 12 different models of pulse feeding were performed to evaluate the effect of pulse feeding at different cultivation phases and pulse frequency on cell growth, acetic acid accumulation, and human‐like collagen (HLC) synthesis. The results showed that the acetate concentration was kept at a low level (below 0.5 g/L) in all cases when pulse feeding was introduced before induction, whereas the pulse frequency affected cytoactivity significantly through cell growth rate, oxygen uptake rate, carbon dioxide evolution rate, and the synthesis of the target protein. The final biomass and HLC reached 75.46 and 7.26 g/L, respectively, in the model of 8‐Sec feedings per 188 Sec. After induction, the pulse frequency had a great effect on HLC synthesis after high‐temperature induction; low frequency was adverse to microorganisms. The model of 3‐Sec feeding per 27 Sec was best and resulted in the highest biomass and HLC production. Compared to the pseudo‐exponential feeding, pulse feeding reduced acetic acid accumulation effectively.