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.
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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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Lin, KH., Chien, MF., Hsieh, JL. et al. Mercury resistance and accumulation in Escherichia coli with cell surface expression of fish metallothionein. Appl Microbiol Biotechnol 87, 561–569 (2010). https://doi.org/10.1007/s00253-010-2466-x
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DOI: https://doi.org/10.1007/s00253-010-2466-x