Genetic engineering modification and fermentation optimization for extracellular production of recombinant proteins using Escherichia coli

  title={Genetic engineering modification and fermentation optimization for extracellular production of recombinant proteins using Escherichia coli},
  author={Yuling Zhou and Zhenghui Lu and Xiang Wang and Jonathan Nimal Selvaraj and Guimin Zhang},
  journal={Applied Microbiology and Biotechnology},
As a common expression host, Escherichia coli has received more and more attention due to the recently developed secretory expression system, which offers advantages like reduced downstream bioprocesses and improved product quality. These advantages, coupled with high-density fermentation technology, make it a preferred system for large-scale production of many proteins utilized in industry and agriculture at a reduced process cost. To improve the secretion efficiency of target proteins… 

Stepwise optimization of recombinant protein production in Escherichia coli utilizing computational and experimental approaches

A stepwise methodology linking the factors from both levels for optimizing the production of soluble recombinant protein in E. coli is proposed, which can facilitate the optimization of gene- and protein-based factors in silico tools.

Inhibition of E. coli Host RNA Polymerase Allows Efficient Extracellular Recombinant Protein Production by Enhancing Outer Membrane Leakiness

It is shown that inducible growth repression in the X‐press strain greatly mitigates the effect of metabolic burden under different process conditions, and demonstrates that the X-press strain constitutes a valuable host for extracellular production of recombinant protein with E. coli.

Triggering outer membrane leakiness of a novel E. coli strain for recombinant protein production

It is demonstrated that the enGenes-X-press strain constitutes a superior host for extracellular production of recombinant protein, and confined the parameter space in which outer membrane leakiness can be controlled, while maintaining cell viability.

Enhancing extracellular production of lipoxygenase in Escherichia coli by signal peptides and autolysis system

The different signal peptides and cell autolysis system were developed and characterized forextracellular LOX production in E. coli and the extracellular production of LOX was achieved and the content of inclusion bodies in the cell was reduced by optimizing cell lysis conditions.

Production of truncated peptide (cellobiohydrolase Cel6A) by Trichoderma reesei expressed in Escherichia coli

This work attempted to use codon optimization to enhance Cel6A expression in Escherichia coli, directing efforts to elucidate the causes of the production of truncated cellulases by this bacterial factory.

A PhoA-STII Based Method for Efficient Extracellular Secretion and Purification of Fab from Escherichia coli.

A protocol based on the alkaline phosphatase (phoA) promoter and the heat-stable enterotoxin II (STII) leader sequence is provided, which could be generally used for the efficient production of Fabs.

Improved extracellular secretion of β-cyclodextrin glycosyltransferase from Escherichia coli by glycine supplementation without apparent cell lysis

An effective glycine feeding strategy could enhance the extracellular secretion of β-CGTase without adverse effects on cell viability, and could be applied potentially to enhance the secretion of a recombinant protein to the culture medium from E. coli.

Genetic Tools and Techniques for Recombinant Expression in Thermophilic Bacillaceae

The biology of thermophilic Bacillaceae is described and genetic tools and methods enabling use of these organisms as hosts for recombinant protein production are focused on.

Enhancement of extracellular bispecific anti‐MUC1 nanobody expression in E. coli BL21 (DE3) by optimization of temperature and carbon sources through an autoinduction condition

To increase both intracellular and extracellular nanobody production, initially, the experiments were performed for the key factors including temperature and duration of protein expression, and the results show less impurity and toxicity in the final product were observed.



Extracellular recombinant protein production from Escherichia coli

This review provides an overview of recent developments in engineering E. coli for extracellular production of recombinant proteins and an analysis of pros and cons of each strategy.

Secretory and extracellular production of recombinant proteins using Escherichia coli

  • J. ChoiS. Y. Lee
  • Biology, Engineering
    Applied Microbiology and Biotechnology
  • 2004
Recent advances in secretory and extracellular production of recombinant proteins using E. coli are discussed, including the twin-arginine translocation system, which has recently been employed for the efficient secretion of folded proteins.

High-throughput recombinant protein expression in Escherichia coli: current status and future perspectives

This review describes several state-of-the-art approaches suitable for the cloning, expression and purification, conducted in parallel, of numerous molecules, and discusses recent progress related to soluble protein expression, mRNA folding, fusion tags, post-translational modification and production of membrane proteins.

An Engineered Survival-Selection Assay for Extracellular Protein Expression Uncovers Hypersecretory Phenotypes in Escherichia coli.

A generalizable survival-based selection strategy is developed that effectively couples extracellular protein secretion to antibiotic resistance and enables facile isolation of rare mutants from very large populations based simply on cell growth and is anticipated to be leveraged to better understand the YebF pathway and other secretory mechanisms.

Enhancement of extracellular pullulanase production from recombinant Escherichia coli by combined strategy involving auto-induction and temperature control

In order to reduce the extended production period, a two-stage temperature control strategy was developed and its application not only reduced the production period from 72 to 36 h, but also further enhanced the yield of extracellular pullulanase.

Recombinant protein secretion in Escherichia coli.