Overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems

Abstract

During the proteomics period, the growth in the use of recombinant proteins has increased greatly in the recent years. Bacterial systems remain most attractive due to low cost, high productivity, and rapid use. However, the rational choice of the adequate promoter system and host for a specific protein of interest remains difficult. This review gives an overview of the most commonly used systems: As hosts, Bacillus brevis, Bacillus megaterium, Bacillus subtilis, Caulobacter crescentus, other strains, and, most importantly, Escherichia coli BL21 and E. coli K12 and their derivatives are presented. On the promoter side, the main features of the l-arabinose inducible araBAD promoter (PBAD), the lac promoter, the l-rhamnose inducible rhaP BAD promoter, the T7 RNA polymerase promoter, the trc and tac promoter, the lambda phage promoter p L , and the anhydrotetracycline-inducible tetA promoter/operator are summarized.

A biological treasure metagenome: pave a way for big science

“…This information results in various fi elds of applications that include, but are not limited to, environmental, ecological and industrial needs, some of which also provide a clue for the origin and minimal genome of living organisms researchers have tried RT-PCR, gene cassette PCR and microarrays for chasing a limited region of whole gene without restricting the sequence space of the other region [44,45]. In addition, the improvement of the vector system and the extent of the host spectrum range are being attempted for the effi cient construction of libraries [46], which are very important in increasing the frequency of fi nding useful genetic resources. These efforts will increase the use of the approach based on gene expression prerequisite to screening.…”

How to approach heterogeneous protein expression for biotechnological use: An overview

“…Therefore, E. coli strains have been developed that lack nucleases and proteases, to prevent degradation of the recombinant DNA and the produced protein. A variety of E. coli strains with various desired properties have been developed by mutations of specific genes (Baneyx 1999;Yokoyama 2003;Klock et al 2005;Terpe 2006). The most common E. coli strains include BL21 (derived from the B strain) and derivatives thereof.…”

A method for the prevention of thrombin-induced degradation of recombinant proteins

“…This method is generally applicable to all recombinant proteins that require the thrombin for the cleavage of affinity tags for purification. To our knowledge, this is the first report of an efficient and reliable method for the separation of residual thrombin from purified recombinant proteins.Keywords recombinant protein; thrombin; heparin; degradation; structure; non-specific Large scale production and purification of pure recombinant proteins is the first important step towards understanding their structure-function relationship [1,2]. In general, recombinant proteins are bioengineered with affinity tag either at their N-or C-terminus [2].…”

“…Large scale production and purification of pure recombinant proteins is the first important step towards understanding their structure-function relationship [1,2]. In general, recombinant proteins are bioengineered with affinity tag either at their N-or C-terminus [2].…”

“…In general, recombinant proteins are bioengineered with affinity tag either at their N-or C-terminus [2]. These affinity tags not only facilitate easy purification of recombinant proteins, but are also known to aid in overexpression of proteins in soluble forms [3].…”