title: Genome engineering for improved recombinant protein expression in Escherichia coli.
abstract: A metabolic engineering perspective which views recombinant protein expression as a multistep pathway allows us to move beyond vector design and identify the downstream rate limiting steps in expression. In E.coli these are typically at the translational level and the supply of precursors in the form of energy, amino acids and nucleotides. Further recombinant protein production triggers a global cellular stress response which feedback inhibits both growth and product formation. Countering this requires a system level analysis followed by a rational host cell engineering to sustain expression for longer time periods. Another strategy to increase protein yields could be to divert the metabolic flux away from biomass formation and towards recombinant protein production. This would require a growth stoppage mechanism which does not affect the metabolic activity of the cell or the transcriptional or translational efficiencies. Finally cells have to be designed for efficient export to prevent buildup of proteins inside the cytoplasm and also simplify downstream processing. The rational and the high throughput strategies that can be used for the construction of such improved host cell platforms for recombinant protein expression is the focus of this review.
authors.affiliation: School of Biotechnology, Jawaharlal Nehru University, New Delhi, 110067, India. shubhashreemahalik@gmail.com.
authors.forename: Shubhashree
authors.initials: S
authors.lastname: Mahalik
authors.affiliation: School of Biotechnology, Jawaharlal Nehru University, New Delhi, 110067, India. ashish.kumar07@gmail.com.
authors.forename: Ashish K
authors.initials: AK
authors.lastname: Sharma
authors.affiliation: School of Biotechnology, Jawaharlal Nehru University, New Delhi, 110067, India. kjmukherjee@mail.jnu.ac.in.
authors.forename: Krishna J
authors.initials: KJ
authors.lastname: Mukherjee
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D016680:Genome, Bacterial; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D011994:Recombinant Proteins