Table 1 Common techniques to optimize recombinant protein production in bacteria

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.