Statistical approaches to maximize recombinant protein expression in Escherichia coli: A general review

A comparison of statistical approaches used for the optimization of soluble protein expression in Escherichia coli

“…Changing the culture conditions to increase the solubility of a recombinant protein is not new, however the optimization of the soluble expression is routinely performed using a one-factor-at-a-time approach with the different expression variables being tested independently from each other [12]. Optimization of soluble protein expression using a traditional one-factor-at-a-time approach is very labor-intensive endeavor because requires a relative large number of experiments and usually fails to anticipate optimal conditions [14,31].…”

“…Although significant improvements and a wide range of systems for M A N U S C R I P T A C C E P T E D ACCEPTED MANUSCRIPT 4 heterologous protein expression in E. coli have been developed, obtaining soluble protein for functional and structural studies still remains a considerable bottleneck [11]. The expression and solubility propensity strongly depend on the target protein, which makes it difficult to deduce a ''consensus approach'' for expression [12]. In general, the production of soluble and active proteins is influenced by several factors including expression host, fusion tag, induction temperature and time.…”

“…The production of recombinant proteins is generally performed using a trial-and-error approach, with the different expression variables being tested independently from each other. Thus, variable interactions are lost which makes the trial-and-error approach timeconsuming [12]. [13].…”

Stepwise optimization of a low-temperature Bacillus subtilis expression system for “difficult to express” proteins

“…Cold-inducible or cold-active promoters can facilitate efficient expression of a target gene at suboptimal growth temperatures (Papaneophytou and Kontopidis 2014;Vasina and Baneyx 1997). Since a P des -regulated cold-inducible expression system for B. subtilis has been published earlier (Thuy Le and Schumann 2007), our study focused on the optimization of this system by additional regulatory sequences, e.g., derived from the B. subtilis major cold-shock gene cspB.…”

Expression, characterization and mutagenesis of an FAD-dependent glucose dehydrogenase from Aspergillus terreus

“…Other hosts such as P. pastoris can be an alternative for production, but expression in E. coli is preferable for the large-scale production required for structural studies and potential industrial applications. Extensive work has been carried out to improve E. coli as a host for recombinant protein expression [21,31]. It is well known that promoter plays an important role in protein heterologous production, and is particularly important for producing functional protein [32].…”