Repetitive Fed-Batch: A Promising Process Mode for Biomanufacturing With E. coli

Area of Science:

• Biotechnology
• Microbial Fermentation
• Protein Expression

Background:

• Industrial recombinant protein production predominantly uses fed-batch cultures in Escherichia coli (E. coli).
• Current methods involve time- and cost-intensive equipment setup and cleaning cycles.
• Continuous biomanufacturing in microbes is limited by productivity variations, and repetitive fed-batch studies are scarce for E. coli.

Purpose of the Study:

• To develop and evaluate a repetitive fed-batch technology for recombinant protein production in E. coli.
• To compare the space-time yield of repetitive fed-batch processes against single-cycle fed-batch and chemostat cultivations.
• To assess the economic and environmental advantages of repetitive fed-batch technology.

Main Methods:

• Development of a mixed feed approach to enable repetitive fed-batch cultivation in E. coli.
• Investigation of cultivation modes including single-cycle fed-batch, two- and three-cycle repetitive fed-batch, and chemostat.
• Utilized two E. coli strains expressing a model protein in the cytoplasm or periplasm.

Main Results:

• Repetitive fed-batch cultivation in E. coli achieved higher space-time yield compared to single-cycle fed-batch.
• The developed repetitive fed-batch process demonstrated potential to outperform continuous biomanufacturing.
• The mixed feeding approach proved suitable for E. coli, improving product throughput and process economics.

Conclusions:

• Repetitive fed-batch technology is highly suitable for recombinant protein production in E. coli.
• This approach optimizes critical equipment utilization, reducing setup and cleaning times.
• The findings suggest a more economical and environmentally favorable biomanufacturing process for E. coli.