Strategies for production of active eukaryotic proteins in bacterial expression system

Microbial β-Glucosidases: Screening, Characterization, Cloning and Applications

“…However, expression in E. coli has several drawbacks such as formation of inclusion bodies, low secretion efficiency, absence of splicing machinery and inability to perform post-translational modifications such as glycosylation explaining why it is not successfully being used for expression of fungal β-glucosidase enzymes [196,197,198]. E.coli is usually transformed with self-replicated plasmid that does not integrate to chromosomal DNA and keeps on replicating itself independently from cell divisions [199,200].…”

Establishment of Stable Chinese Hamster Ovary Cell Line Capable of Expressing Human Recombinant Hemopexin: A Promising Therapeutic Modality Against Hemolytic Anemia

“…Their initial attempts to express human HPX in an E. coli system produced a non-glycosylated protein with no heme-binding ability (15). Protein expression in prokaryotic system is associated with challenges such as limited eukaryotic post-translational machinery function, protein aggregation, and generation of inclusion bodies (33). Therefore, re-solubilization and refolding of recombinant polypeptide chains are required, leading to reduced quality and unexpected biological activity of the final proteins (34).…”

Single cell-level detection and quantitation of leaky protein expression from any strongly regulated bacterial system

“…Many factors favor the use of E. coli as a cell factory. These include its ease of genetic manipulation, its short generation time, the general ease of protein recovery and scale-up from E. coli, and the low costs of growth media and downstream processing [1,3].…”

“…A key problem in heterologous protein expression in E. coli is that certain proteins are toxic even when levels of expression are very low [3,4]. Thus, many efforts have been directed towards the design of tightly regulated, plasmid-borne, bacterial expression systems in which expression only occurs on ''induction'' through addition of an inducer molecule supplied to a growing culture of host bacteria, but not ''prematurely,'' i.e., not before addition of the inducer [5,6].…”

“…Many factors favor the use of E. coli as a cell factory. These include its ease of genetic manipulation, its short generation time, the general ease of protein recovery and scale-up from E. coli, and the low costs of growth media and downstream processing [1,3].A key problem in heterologous protein expression in E. coli is that certain proteins are toxic even when levels of expression are very low [3,4]. Thus, many efforts have been directed towards the design of tightly regulated, plasmid-borne, bacterial expression systems in which expression only occurs on ''induction'' through addition of an inducer molecule supplied to a growing culture of host bacteria, but not ''prematurely,'' i.e., not before addition of the inducer [5,6].…”