Recombinant Protein Production in Escherichia coli: Optimization Strategies

Abstract

The process of making recombinant proteins in Escherichia coli (E. coli) is now commonly used to make a lot of proteins that are used in medicine and industry.  This bacteria, E. coli, is very important in biotechnology because it can quickly and cheaply make foreign proteins. It is used in medicine, diagnosis, and study.  However, making recombinant proteins in E. coli is still a difficult process that is often slowed down by issues like protein solubility, protein clumping, and making sure the recombinant protein folds correctly.  To improve the yield and usefulness of the protein, it is important to make the translation system work better. This paper is mainly about the different methods used to make recombinant protein production in E. coli better.  Some important factors for optimization are choosing the right host strains, expression vectors, and promoters, all of which have a big impact on the protein output and expression levels.  Genes that have been codon-optimized and media that is high in nutrients can both help raise translation levels even more.  Controlling the temperature, the time of the induction, and the quantity of the inducers are also important to keep protein breakdown to a minimum and speed up the folding process. Taking care of problems with protein solubility is another important part of optimization.  For functional studies, soluble proteins work best, but a lot of transgenic proteins tend to form inclusion bodies, which make handling later on harder.  Some ways to stop or lessen the formation of inclusion bodies are to use co-expression systems with chaperones or fusion tags, improve the growth conditions, and change the temperature during induction.  It is also possible to make protein shape and function much better by designing expression vectors that include release signals and post-translational changes. Synthetic biology and metabolic engineering have also led to the creation of stronger E. coli strains that can make proteins with complex modifications, like disulphide bonds or glycosylation, which are often needed for therapeutic proteins to work biologically. It's easier to find and fine-tune the best settings for recombinant protein translation now that high-throughput screening methods are used.