Development of a Robust Pipeline for Maximizing Recombinant Protein Production Efficiency

The main objective of this project is to create a resilient pipeline that optimizes the generation of these hard-to-obtain enzymes using an integrated strategy encompassing bioinformatics, molecular biology, and bioprocess engineering. The project will employ a three-stage approach. In the initial stage, an advanced codon-matching algorithm, recently developed and validated, will be employed to enhance the folding of non-native proteins in E. coli by preserving rare codons at crucial mRNA positions. In the second stage, for proteins that form inclusion bodies or exhibit toxicity, the system will incorporate chaperones, T7 lysozyme, and extracellular expression. Finally, a continuous fermentation process will be established to boost productivity and cut down on costs.