Recombinant protein production in bacterial hosts.

The production of recombinant proteins is crucial for both the development of new protein drugs and the structural determination of drug targets. As such, recombinant protein production has a major role in drug development. Bacterial hosts are commonly used for the production of recombinant proteins, accounting for approximately 30% of current biopharmaceuticals on the market. In this review, I introduce fundamental concepts in recombinant protein production in bacteria, from drug development to production scales. Recombinant protein production processes can often fail, but how can this failure be minimised to rapidly deliver maximum yields of high-quality protein and so accelerate drug discovery?

[1]  K. Terpe Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems , 2002, Applied Microbiology and Biotechnology.

[2]  C. Elvin,et al.  Modified bacteriophage lambda promoter vectors for overproduction of proteins in Escherichia coli. , 1990, Gene.

[3]  Alan J Wolfe,et al.  Glucose metabolism at high density growth of E. coli B and E. coli K: differences in metabolic pathways are responsible for efficient glucose utilization in E. coli B as determined by microarrays and Northern blot analyses. , 2005, Biotechnology and bioengineering.

[4]  Arti Kapi The evolving threat of antimicrobial resistance: Options for action , 2014 .

[5]  U. Rinas,et al.  Side effects of chaperone gene co-expression in recombinant protein production , 2010, Microbial cell factories.

[6]  K. Terpe Overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems , 2006, Applied Microbiology and Biotechnology.

[7]  A. Riggs,et al.  Expression in Escherichia coli of a chemically synthesized gene for the hormone somatostatin. , 1977, Science.

[8]  S. Singh,et al.  Solubilization and refolding of bacterial inclusion body proteins. , 2005, Journal of bioscience and bioengineering.

[9]  J. Walker,et al.  Over-production of proteins in Escherichia coli: mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels. , 1996, Journal of molecular biology.

[10]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[11]  Michael Sauer,et al.  Recombinant protein production in yeasts. , 2012, Methods in molecular biology.

[12]  R. Lenski,et al.  Tracing ancestors and relatives of Escherichia coli B, and the derivation of B strains REL606 and BL21(DE3). , 2009, Journal of molecular biology.

[13]  R. Neutze,et al.  Effective high-throughput overproduction of membrane proteins in Escherichia coli. , 2008, Protein expression and purification.

[14]  A. N. Weir,et al.  Expression of antibody fragments by periplasmic secretion in Escherichia coli. , 2005, Methods in molecular biology.

[15]  H. Nothaft,et al.  Bacterial Protein N-Glycosylation: New Perspectives and Applications* , 2013, The Journal of Biological Chemistry.

[16]  R. Freedman,et al.  High‐yield export of a native heterologous protein to the periplasm by the tat translocation pathway in Escherichia coli , 2012, Biotechnology and bioengineering.

[17]  N. Majdalani,et al.  The RpoS-mediated general stress response in Escherichia coli. , 2011, Annual review of microbiology.

[18]  P. Langella,et al.  Lactococcus lactis, an Efficient Cell Factory for Recombinant Protein Production and Secretion , 2007, Journal of Molecular Microbiology and Biotechnology.

[19]  Samuel Wagner,et al.  Tuning Escherichia coli for membrane protein overexpression , 2008, Proceedings of the National Academy of Sciences.

[20]  George Georgiou,et al.  Preparative expression of secreted proteins in bacteria: status report and future prospects. , 2005, Current opinion in biotechnology.

[21]  Antonio Villaverde,et al.  Localization of Functional Polypeptides in Bacterial Inclusion Bodies , 2006, Applied and Environmental Microbiology.

[22]  Frank Hoffmann,et al.  Stress induced by recombinant protein production in Escherichia coli. , 2004, Advances in biochemical engineering/biotechnology.

[23]  I. Henderson,et al.  A generalised module for the selective extracellular accumulation of recombinant proteins , 2012, Microbial Cell Factories.

[24]  Gyun Min Lee,et al.  CHO cells in biotechnology for production of recombinant proteins: current state and further potential , 2012, Applied Microbiology and Biotechnology.

[25]  B. Gronenborn Overproduction of phage Lambda repressor under control of the lac promotor of Escherichia coli , 1976, Molecular and General Genetics MGG.

[26]  G. Guiochon,et al.  Separation science is the key to successful biopharmaceuticals. , 2011, Journal of chromatography. A.

[27]  S. Lee,et al.  High cell-density culture of Escherichia coli. , 1996, Trends in biotechnology.

[28]  N. W. Davis,et al.  The complete genome sequence of Escherichia coli K-12. , 1997, Science.

[29]  D. Sherratt,et al.  Escherichia coli strains that allow antibiotic-free plasmid selection and maintenance by repressor titration. , 2001, Nucleic acids research.

[30]  Tim W. Overton,et al.  Exploitation of GFP fusion proteins and stress avoidance as a generic strategy for the production of high-quality recombinant proteins. , 2009, FEMS microbiology letters.

[31]  Gary Walsh,et al.  Biopharmaceutical benchmarks 2010 , 2010, Nature Biotechnology.

[32]  J. Brosius,et al.  Spacing of the -10 and -35 regions in the tac promoter. Effect on its in vivo activity. , 1985, The Journal of biological chemistry.

[33]  I. S. Johnson Human insulin from recombinant DNA technology. , 1983, Science.

[34]  D. Waugh,et al.  Making the most of affinity tags. , 2005, Trends in biotechnology.

[35]  F. Baneyx,et al.  Recombinant protein folding and misfolding in Escherichia coli , 2004, Nature Biotechnology.

[36]  Rebecca Page,et al.  Strategies to maximize heterologous protein expression in Escherichia coli with minimal cost. , 2007, Protein expression and purification.

[37]  Alois Jungbauer,et al.  Protein Chromatography: Process Development and Scale-Up , 2010 .

[38]  C. Harwood,et al.  Heterologous protein secretion by bacillus species from the cradle to the grave. , 2010, Advances in applied microbiology.

[39]  A. P. Chapman,et al.  PEGylated antibodies and antibody fragments for improved therapy: a review. , 2002, Advanced drug delivery reviews.

[40]  J. Collet,et al.  Oxidative protein folding in bacteria , 2002, Molecular microbiology.

[41]  I. Henderson,et al.  Type V Protein Secretion Pathway: the Autotransporter Story , 2004, Microbiology and Molecular Biology Reviews.

[42]  Gary Walsh,et al.  Post-translational modifications of protein biopharmaceuticals. , 2009, Drug discovery today.

[43]  A. de Marco,et al.  Microbial Cell Factories Strategies for Successful Recombinant Expression of Disulfide Bond-dependent Proteins in Escherichia Coli , 2022 .

[44]  D. Summers,et al.  Recombinant protein secretion in Escherichia coli. , 2005, Biotechnology advances.

[45]  Chung-Jr Huang,et al.  Industrial production of recombinant therapeutics in Escherichia coli and its recent advancements , 2012, Journal of Industrial Microbiology & Biotechnology.

[46]  R. Sodoyer,et al.  Antibiotic-free selection in E. coli: new considerations for optimal design and improved production , 2010, Microbial cell factories.

[47]  S. Valla,et al.  Positively regulated bacterial expression systems , 2008, Microbial biotechnology.

[48]  Sarah E. Ades,et al.  Regulation by destruction: design of the sigmaE envelope stress response. , 2008, Current opinion in microbiology.

[49]  F. Bontems,et al.  Expression of highly toxic genes in E. coli: special strategies and genetic tools. , 2006, Current protein & peptide science.

[50]  Takashi Yura,et al.  Convergence of Molecular, Modeling, and Systems Approaches for an Understanding of the Escherichia coli Heat Shock Response , 2008, Microbiology and Molecular Biology Reviews.

[51]  Daniel G Bracewell,et al.  Advances in product release strategies and impact on bioprocess design. , 2009, Trends in biotechnology.

[52]  G. Petersen,et al.  Current strategies for the use of affinity tags and tag removal for the purification of recombinant proteins. , 2006, Protein expression and purification.

[53]  D. Belin,et al.  Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter , 1995, Journal of bacteriology.

[54]  M. Hyvönen,et al.  T7 vectors with modified T7lac promoter for expression of proteins in Escherichia coli. , 1996, Analytical biochemistry.

[55]  T. Ng,et al.  Immobilized metal ion affinity chromatography: a review on its applications , 2012, Applied Microbiology and Biotechnology.

[56]  A. Driessen,et al.  Sec- and Tat-mediated protein secretion across the bacterial cytoplasmic membrane--distinct translocases and mechanisms. , 2008, Biochimica et biophysica acta.

[57]  M. Day,et al.  The biology of plasmids. , 1987, Science progress.

[58]  F. Studier Use of bacteriophage T7 lysozyme to improve an inducible T7 expression system. , 1991, Journal of molecular biology.

[59]  W. Reznikoff,et al.  The lactose operon‐controlling elements: a complex paradigm , 1992, Molecular microbiology.

[60]  Rachel Chen Bacterial expression systems for recombinant protein production: E. coli and beyond. , 2012, Biotechnology advances.

[61]  Esther Vázquez,et al.  Microbial factories for recombinant pharmaceuticals , 2009 .

[62]  Gary Walsh,et al.  Biopharmaceutical benchmarks , 2000, Nature Biotechnology.

[63]  M. De Mey,et al.  Increasing recombinant protein production in Escherichia coli K12 through metabolic engineering. , 2013, New biotechnology.

[64]  R. Hall,et al.  Use of GFP fusions for the isolation of Escherichia coli strains for improved production of different target recombinant proteins. , 2011, Journal of biotechnology.

[65]  E. Keshavarz‐Moore,et al.  Development of a simple method for the recovery of recombinant proteins from the Escherichia coli periplasm , 1996 .

[66]  A. Villaverde,et al.  The conformational quality of insoluble recombinant proteins is enhanced at low growth temperatures , 2007, Biotechnology and bioengineering.

[67]  Udo Oppermann,et al.  Codon optimization can improve expression of human genes in Escherichia coli: A multi-gene study. , 2008, Protein expression and purification.

[68]  D. Jin,et al.  Growth rate regulation in Escherichia coli. , 2012, FEMS microbiology reviews.

[69]  H. P. Sørensen,et al.  Soluble expression of recombinant proteins in the cytoplasm of Escherichia coli , 2005 .

[70]  Viktor Menart,et al.  Production of Nonclassical Inclusion Bodies from Which Correctly Folded Protein Can Be Extracted , 2008, Biotechnology progress.

[71]  Gary Walsh,et al.  Biopharmaceutical benchmarks 2014 , 2014, Nature Biotechnology.

[72]  Joseph Shiloach,et al.  Growing E. coli to high cell density--a historical perspective on method development. , 2005, Biotechnology advances.

[73]  J. Beckwith,et al.  Disulfide bond formation in the Escherichia coli cytoplasm: an in vivo role reversal for the thioredoxins , 1998, The EMBO journal.