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

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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.

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Acknowledgement

This study was supported by grants from Council of Scientific and Industrial Research (CSIR), India.

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Authors and Affiliations

  1. Centre for Biotechnology, Anna University, Chennai, 600025, India

    K. Senthil & P. Gautam

Authors
  1. K. Senthil
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  2. P. Gautam
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Correspondence to P. Gautam.

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Purpose of work: The objective of this work was to overexpress mercury transporter (merT) protein and to circumvent the expression induced toxicity in E. coli. We further investigated the optimum detergents and solubility conditions required for single-step purification of merT by immobilized metal affinity chromatography.

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Senthil, K., Gautam, P. Expression and single-step purification of mercury transporter (merT) from Cupriavidus metallidurans in E. coli . Biotechnol Lett 32, 1663–1666 (2010). https://doi.org/10.1007/s10529-010-0337-2

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  • DOI: https://doi.org/10.1007/s10529-010-0337-2

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