Alternative production process strategies in E. coli improving protein quality and downstream yields

Influence of growth temperature on the production of antibody Fab fragments in different microbes: A host comparative analysis

“…Reduction of the cultivation temperature had a positive effect on product yield of a Fab 3H6 in E. coli , P. pastoris , and T. reesei , whereas no significant effect on product yield was observed in S. cerevisiae . For prokaryotic hosts, most notably E. coli , it is well established that reduced temperature favors recombinant protein production (1, 17, 18 and Table 1); however, the majority of these data was acquired in batch culture with variable specific growth rates. Consequently, these reports also suggest that reduced growth rate at decreased temperature is responsible for increased productivity, resulting in lower competition for metabolites and precursor substances of recombinant proteins and host cell metabolism.…”

“…Examples range from bacteria to yeasts and higher eukaryotes, especially mammalian cells (see Table 1). For bacterial production systems, the positive effect of lower growth temperature is usually attributed to facilitated protein folding, and consequently higher yields of correctly folded soluble product 1, 17, 18. Mammalian cells react to reduced cultivation temperature (e.g., reduction from 37°C to less than 30°C) with an extended period of viability, which is leading to higher product yield (reviewed by19).…”

Simulation and prediction of protein production in fed‐batch E. coli cultures: An engineering approach

“…Biomass concentration at induction time ranged from 8 to 47 gDCW · L −1 , being the later the maximum allowing a final biomass concentration compatible with the aeration capacity of the fermentor. Finally, specific growth rate ranged from 0.1 to 0.22 h −1 , avoiding too slow processes and excessive acetic acid production (Ruiz et al, ).…”

Process optimization of high-level extracellular production of alkaline pectate lyase in recombinant Escherichia coli BL21 (DE3)

“…In order to achieve HCDC of E. coli, a three-step glycerol feeding strategy was employed in our study. In the first step, exponential feeding was carried out and acetate formation was prevented by controlling the specific growth rate of the cells at 0.25 h −1 (below the threshold of the growth rate for acetate formation, which was ∼0.32 h −1 ) [10,15,21,27]. After that, when the feeding rate reached 12.5 mL h −1 L −1 which was the maximal feeding rate for avoiding acetate formation under the maximum aerobic conditions used in this study, the second-step feeding started and kept this feeding rate.…”