Increasing recombinant protein production in Escherichia coli through metabolic and genetic engineering

• Published in: Journal of industrial microbiology & biotechnology Vol. 38; no. 12; pp. 1891 - 1910
• Main Authors: Waegeman, Hendrik, Soetaert, Wim
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.12.2011; Springer; Oxford University Press
• Subjects: acetates; Bacillus subtilis; bacteria; Biochemistry; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; biopharmaceuticals; Biotechnology; Chromosomes; Cloning; E coli; Enzymes; Escherichia coli; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli - metabolism; eukaryotic cells; Fundamental and applied biological sciences. Psychology; Genes; Genetic Engineering; Genomes; hosts; Inorganic Chemistry; insects; Life Sciences; Metabolism; Microbiology; momentum; Pharmaceuticals; Pichia pastoris; Plasmids; post-translational modification; Protein Processing, Post-Translational; Proteins; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Review; RNA polymerase; Saccharomyces cerevisiae; Studies; Vectors (Biology); Europe; Industrial enzyme; Recombinant protein production; Engineering; Heterologous protein; Biopharmaceutical; Drug; Enzyme; Escherichia coli; Biopharmaceuticals; Production; Bacteria; Metabolic engineering; Genetic engineering; Recombinant protein; Enterobacteriaceae
• ISSN: 1367-5435; 1476-5535
• DOI: 10.1007/s10295-011-1034-4

Different hosts have been used for recombinant protein production, ranging from simple bacteria, such as Escherichia coli and Bacillus subtilis, to more advanced eukaryotes as Saccharomyces cerevisiae and Pichia pastoris, to very complex insect and animal cells. All have their advantages and drawbacks and not one seems to be the perfect host for all purposes. In this review we compare the characteristics of all hosts used in commercial applications of recombinant protein production, both in the area of biopharmaceuticals and industrial enzymes. Although the bacterium E. coli remains a very often used organism, several drawbacks limit its possibility to be the first-choice host. Furthermore, we show what E. coli strains are typically used in high cell density cultivations and compare their genetic and physiological differences. In addition, we summarize the research efforts that have been done to improve yields of heterologous protein in E. coli, to reduce acetate formation, to secrete the recombinant protein into the periplasm or extracellular milieu, and to perform post-translational modifications. We conclude that great progress has been made in the incorporation of eukaryotic features into E. coli, which might allow the bacterium to regain its first-choice status, on the condition that these research efforts continue to gain momentum.