Understanding the Art of Producing Protein and Nonprotein Molecules in Escherichia coli
- 1 January 2001
- journal article
- review article
- Published by Springer Science and Business Media LLC in Molecular Biotechnology
- Vol. 19 (3), 251-268
- https://doi.org/10.1385/mb:19:3:251
Abstract
The high-level production of functional proteins in E. coli is a very extense field of research in biotechnology. A number of important aspects to be considered in the initial design of an expression system and their interplay, were clear years ago. However, in recent times, strategies that go beyond transcription, translation, stability, vector, and strain choice, have been developed; so now expression of active peptides can be viewed as a more integrated process. Coexpression of protein subunits, foldases and chaperones, protein folding, location and purification schemes, metabolic engineering of the cell’s central metabolism, and in vitro refolding strategies, are some of the novelties that are now available to aid in the success of an efficient expression system for active heterologous proteins. This review presents a compilation of the basic issues that influence the success in the production of protein and nonprotein products in Escherichia coli, as well as some general strategies designed to facilitate downstream process operations and improve biosynthesis yields.Keywords
This publication has 93 references indexed in Scilit:
- Theory for the Systemic Definition of Metabolic Pathways and their use in Interpreting Metabolic Function from a Pathway-Oriented PerspectiveJournal of Theoretical Biology, 2000
- Prokaryotic Gene Fusion Expression Systems and their Use in Structural and Functional Studies of ProteinsPreparative Biochemistry & Biotechnology, 1999
- Participation of IHF and a distant UP element in the stimulation of the phage λ PLpromoterMolecular Microbiology, 1998
- Affinity Fusion Strategies for Detection, Purification, and Immobilization of Recombinant ProteinsProtein Expression and Purification, 1997
- Growth rate-optimised tRNA abundance and codon usageJournal of Molecular Biology, 1997
- Use of a Modified Tryptophanase Promoter to Direct High-Level Expression of Foreign Proteins in E. coliAnnals of the New York Academy of Sciences, 1996
- A chromosomal expression vector for Escherichia coli based on the bacteriophage MuGene, 1993
- Culture conditions differentially affect the translation of individual Escherichia coli mRNAsJournal of Molecular Biology, 1992
- Control of in vivo proteolysis in the production of recombinant proteinsTrends in Biotechnology, 1992
- Ribosomal RNA operon anti-terminationJournal of Molecular Biology, 1989