Expression and single-step purification of mercury transporter (merT) from Cupriavidus metallidurans in E. coli

Abstract

The mercury transporter, merT, from Cupriavidus metallidurans was cloned into pRSET-C and expressed in various E. coli hosts. Expression of merT gene failed in common expression hosts like E. coli BL21(DE3), E. coli BL21(DE3)pLysS and E. coli GJ1158 due to expression induced toxicity. The protein was successfully expressed in E. coli C43(DE3) as inclusion bodies. The inclusion bodies were solubilized with Triton X-100 detergent. The detergent solubilized protein with N-terminal His-tag was purified in a single-step by immobilized metal affinity chromatography with a yield of 8 mg l(-1).

Complete Genome Sequence of a Marine Bacterium, Pseudomonas pseudoalcaligenes Strain S1, with High Mercury Resistance and Bioaccumulation Capacity

“…Genes for mercuric regulation ( merR ), mercuric transport and binding ( merT and merP ), and metal ion efflux pump (CzcA) were identified from the P. pseudoalcaligenes S1 complete genome. They may be responsible for the adaptation of mercury contamination through transmembrane transportation and active efflux ( 10 – 13 ), therefore, they may help practice the function of high mercury resistance and accumulation in the marine bacterium P. pseudoalcaligenes S1. In addition, the gene for mercuric reduction ( merA ) was also predicted from our transcriptome data; however, its function remains a mystery.…”

Heterologous Soluble Expression of Recombinant OmpR of <i>Aeromonas hydrophila</i> and Its Immunogenic Potential

“…Since the OmpR homologue is present in E. coli, the protein is native to the host cell and hence leaky expression of the rOmpR would not adversely affect the cellular metabolism of the host cell. Unlike other membrane protein, MerT, which could not be expressed in GJ1158 cells due to expression induced toxicity [31], OmpR of A. hydrophila could be successfully expressed in these E. coli host cells. Use of NaCl as an inducer has an advantage over IPTG with respect to its low cost and relatively no toxicity.…”

Improving the mda-7/IL-24 refolding and purification process using optimized culture conditions based on the structure characteristics of inclusion bodies

“…Thus, the structural characteristics of IBs could affect their solubilization efficiency [19]. There were many studies about the refolding methods of IBs in vitro [20][21][22]. However, no previous studies have attempted to improve the structure of mda-7/IL-24 IBs by optimizing the culture conditions to facilitate the subsequent IB renaturation and purification process.…”