Abstract
While the nutrient limited fed-batch technology is the standard of the cultivation of microorganisms and production of heterologous proteins in industry, despite its advantages in view of metabolic control and high cell density growth, shaken batch cultures are still the standard for protein production and expression screening in molecular biology and biochemistry laboratories. This is due to the difficulty and expenses to apply a controlled continuous glucose feed to shaken cultures. New ready-made growth media, e.g. by biocatalytic release of glucose from a polymer, offer a simple solution for the application of the fed-batch principle in shaken plate and flask cultures. Their wider use has shown that the controlled diet not only provides a solution to obtain significantly higher cell yields, but also in many cases folding of the target protein is improved by the applied lower growth rates; i.e. final volumetric yields for the active protein can be a multiple of what is obtained in complex medium cultures. The combination of the conventional optimization approaches with new and easy applicable growth systems has revolutionized recombinant protein production in Escherichia coli in view of product yield, culture robustness as well as significantly increased cell densities. This technical development establishes the basis for successful miniaturization and parallelization which is now an important tool for synthetic biology and protein engineering approaches. This review provides an overview of the recent developments, results and applications of advanced growth systems which use a controlled glucose release as substrate supply.
Highlights
For many reasons Escherichia coli is still the preferred choice as a host system for protein production
The preexpression of Erv1p sulfhydryl oxidase and disulfide bond isomerases for example is a robust technique for the production of disulfide bonds containing proteins [6, 7]
Growth systems that can offer controlled cultivation conditions can be used as an easy tool for the production of a variety of proteins in E. coli—usually E. coli BL21 (DE3)
Summary
For many reasons Escherichia coli is still the preferred choice as a host system for protein production. Growth systems that can offer controlled cultivation conditions can be used as an easy tool for the production of a variety of proteins in E. coli—usually E. coli BL21 (DE3).
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