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Applied Microbiology and Biotechnology

, Volume 87, Issue 2, pp 561–569 | Cite as

Mercury resistance and accumulation in Escherichia coli with cell surface expression of fish metallothionein

  • Kuo-Hsing Lin
  • Mei-Fang Chien
  • Ju-Liang Hsieh
  • Chieh-Chen HuangEmail author
Biotechnologically Relevant Enzymes and Proteins

Abstract

Recombinant tilapia (Oreochromis mossambicus) fish metallothionein (MT) was used as a surface biosorbent for mercury removal in Escherichia coli. Fish MT conferred better resistance than did mouse or human MT. When tilapia MT (tMT) was fused with an outer-membrane protein, outer membrane protein C (OmpC), the membrane-targeted fusion protein, OmpC–tMT, gave enhanced resistance compared with cytoplasmic tMT expressed in the same host cell. The cytoplasmically expressed tMT showed high mercury adsorption (4.3 ± 0.4 mg/g cell dry weight). The cell surface that expressed E. coli showed about 25% higher adsorption ability (5.6 ± 0.4 mg/g) than the cells expressing cytoplasmic MT, attaining almost twice the level of adsorption of the control plasmid (3.0 ± 0.4 mg/g). As MTs are also known for their ability to scavenge hydroxyl-free radicals, it was also shown that tMT exhibited better radical-scavenging activities than glutathione. These results suggest that fish MT has potential for the development of a bioremediation system for mercury removal that protects the harboring E. coli host by free-radical scavenging.

Keywords

Metallothionein Mercury Bioadsorption OmpC Tilapia 

Notes

Acknowledgements

We thank Professor Jo-Shu Chang (National Cheng Kung University, Taiwan) for his kind gift of the pMAL-p2X-MT1A and pMBP-TEV-MT1 plasmids containing human MT1A and mouse MT1 cDNA. We also thank Prof. Su-Mei Wu (National Chia Yi University, Taiwan) for antibody of the rabbit anti-tMT IgG. This work was supported by the grants (NSC 96-3114-P-001-004-Y and NSC 97-3114-P-001-001) from the National Science Council, Taiwan, People’s Republic of China.

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Kuo-Hsing Lin
    • 1
  • Mei-Fang Chien
    • 2
  • Ju-Liang Hsieh
    • 3
  • Chieh-Chen Huang
    • 1
    Email author
  1. 1.Department of Life SciencesNational Chung Hsing UniversityTaichungTaiwan
  2. 2.Center for Marine Environmental StudiesEhime UniversityMatsuyamaJapan
  3. 3.Division of Molecular and Genomic MedicineNational Health Research InstituteMiaoliTaiwan

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