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Increased metal tolerance and bioaccumulation of zinc and cadmium in Chlamydomonas reinhardtii expressing a AtHMA4 C-terminal domain protein

Aniefon Ibuot, Rachel E. Webster, Lorraine E. Williams, View ORCID ProfileJon K. Pittman
doi: https://doi.org/10.1101/2020.02.13.948307
Aniefon Ibuot
aDepartment of Science Technology, Akwa Ibom State Polytechnic, Ikot Osurua, P.M.B 1200.Ikot Ekpene, Akwa Ibom State, Nigeria
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Rachel E. Webster
bThe Manchester Museum, The University of Manchester, Oxford Road, Manchester M13 9PL, UK
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Lorraine E. Williams
cSchool of Biological Sciences, University of Southampton, Building 85, Highfield, Southampton SO17 1BJ, UK
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Jon K. Pittman
dDepartment of Earth and Environmental Sciences, School of Natural Sciences, The University of Manchester, Michael Smith Building, Oxford Road, Manchester M13 9PT, UK
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  • ORCID record for Jon K. Pittman
  • For correspondence: jon.pittman@manchester.ac.uk
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Abstract

The use of microalgal biomass for metal pollutant bioremediation might be improved by genetic engineering to modify the selectivity or capacity of metal biosorption. A plant cadmium (Cd) and zinc (Zn) transporter (AtHMA4) was used as a transgene to increase the ability of Chlamydomonas reinhardtii to tolerate 0.2 mM Cd and 0.3 mM Zn exposure. The transgenic cells showed increased accumulation and internalisation of both metals compared to wild type. AtHMA4 was expressed either as the full-length protein or just the C-terminal tail, which is known to have metal binding sites. Similar Cd and Zn tolerance and accumulation was observed with expression of either the full-length protein or C-terminal domain, suggesting that enhanced metal tolerance was mainly due to increased metal binding rather than metal transport. The effectiveness of the transgenic cells was further examined by immobilisation in calcium alginate to generate microalgal beads that could be added to a metal contaminated solution. Immobilisation maintained metal tolerance, while AtHMA4-expressing cells in alginate showed a concentration-dependent increase in metal biosorption that was significantly greater than alginate beads composed of wild type cells. This demonstrates that expressing AtHMA4 full-length or C-terminus has great potential as a strategy for bioremediation using microalgal biomass.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted June 24, 2020.
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Increased metal tolerance and bioaccumulation of zinc and cadmium in Chlamydomonas reinhardtii expressing a AtHMA4 C-terminal domain protein
Aniefon Ibuot, Rachel E. Webster, Lorraine E. Williams, Jon K. Pittman
bioRxiv 2020.02.13.948307; doi: https://doi.org/10.1101/2020.02.13.948307
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Increased metal tolerance and bioaccumulation of zinc and cadmium in Chlamydomonas reinhardtii expressing a AtHMA4 C-terminal domain protein
Aniefon Ibuot, Rachel E. Webster, Lorraine E. Williams, Jon K. Pittman
bioRxiv 2020.02.13.948307; doi: https://doi.org/10.1101/2020.02.13.948307

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