Genome engineering for improved recombinantprotein expression in Escherichia coli

[摘要] A metabolic engineering perspective which views recombinant proteinexpression as a multistep pathway allows us to move beyond vector design andidentify the downstream rate limiting steps in expression. In E.coli these are typically at the translational leveland the supply of precursors in the form of energy, amino acids and nucleotides.Further recombinant protein production triggers a global cellular stress responsewhich feedback inhibits both growth and product formation. Countering this requiresa system level analysis followed by a rational host cell engineering to sustainexpression for longer time periods. Another strategy to increase protein yieldscould be to divert the metabolic flux away from biomass formation and towardsrecombinant protein production. This would require a growth stoppage mechanism whichdoes not affect the metabolic activity of the cell or the transcriptional ortranslational efficiencies. Finally cells have to be designed for efficient exportto prevent buildup of proteins inside the cytoplasm and also simplify downstreamprocessing. The rational and the high throughput strategies that can be used for theconstruction of such improved host cell platforms for recombinant protein expressionis the focus of this review.

[发布日期] 2014-12-19

[关键词] Recombinant protein expression;Escherichia coli;Metabolic engineering;Genome engineering