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Table 1 Common techniques to optimize recombinant protein production in bacteria

From: A synthetic biology approach to self-regulatory recombinant protein production in Escherichia coli

Method Characteristics
Host strain Natural and engineered host strains can accommodate higher recombinant protein yields
Plasmid copy number The choice of the plasmid backbone influences the production process through gene dosage
Inducer concentration Inducer concentration influences transcription rate and therefore product formation/aggregation rate.
Promoter Different promoters can be considered. Weak/Strong, constitutive and inducible promoters.
Ribosome binding site (RBS) Position and sequence of the RBS influences translational efficiency
mRNA stability and structure mRNA turnover influences the production process as well as mRNA structure can influence ribosome binding and translational efficiency
Codon optimization Codon usage in the sequence of the recombinant gene greatly impacts translation efficiency
Process conditions Temperature, oxygenation, pH and medium osmolarity impact on the production process
Medium composition Optimization of the growth medium can lead to increase of the product yield and decrease of by-product formation
Heat shock protein co-overexpression and knockouts Increased or decreased amount of several molecular chaperones, foldases and proteases influence protein yield and quality.
  1. In depth review of these methods is available in [14]