Production of Soluble Recombinant Proteins in Bacteria

Making the most of affinity tags

“…The most common method that is used to improve the solubility of recombinant proteins in E. coli is to reduce the temperature at which the target protein is being produced, although this is not always effective [7]. However, for some time now it has been recognized that certain affinity tags have the ability to promote the solubility of their fusion partners [8].…”

Expression of catalytically active radish 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase in Escherichia coli

“…As an approach to overcoming the insolubility of HMGR45 and HMGR63, we decreased the temperature of induction, because it has been described that the temperature-dependent reduction of the rate of protein synthesis usually results in an enhanced yield of the soluble expressed proteins [22].…”

“…Nevertheless, we hope that HMGR63 can be recovered in an enzymatically active form after its solubilization and/or reconstitution into its active conformation, which is under current study. Although there exist several factors that account for the insolubility of foreign proteins overexpressed in E. coli [22] specific case the behavior of HMGR63 could presumably be attributed to the presence of the two highly hydrophobic sequences within the membrane domain [6,9]. In the case of cells harboring pHMGR45, HMGR activity increased when the induction was performed at temperatures below 37°C (Fig.…”

A single step purification for recombinant proteins Characterization of a microtubule associated protein (MAP 2) fragment which associates with the type II cAMP‐dependent protein kinase

“…Only in one case, the expression of a 107 amino acid fragment from the RII-anchoring protein, Ht 21, was all of the protein present in inclusion bodies. Formation of inclusion bodies may be decreased if bacteria are induced at 2%30°C or in the presence of 'compatible osmolytes' such as glycyl betaine, which arc believed to cause preferential hydration and stabilization of the recombinant protein structure [26,27]. …”