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Engineered nickel bioaccumulation in Escherichia coli by NikABCDE transporter and metallothionein overexpression

View ORCID ProfileP. Diep, H. Shen, J. A. Wiesner, N. Mykytczuk, V. Papangelakis, A. F. Yakunin, R. Mahadevan
doi: https://doi.org/10.1101/2022.08.26.505507
P. Diep
1BioZone – Centre for Applied Bioscience and Bioengineering, Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON, Canada
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  • ORCID record for P. Diep
H. Shen
1BioZone – Centre for Applied Bioscience and Bioengineering, Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON, Canada
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J. A. Wiesner
2MIRARCO Mining Innovation, Laurentian University, Sudbury ON, Canada
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N. Mykytczuk
2MIRARCO Mining Innovation, Laurentian University, Sudbury ON, Canada
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V. Papangelakis
1BioZone – Centre for Applied Bioscience and Bioengineering, Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON, Canada
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A. F. Yakunin
1BioZone – Centre for Applied Bioscience and Bioengineering, Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON, Canada
3Centre for Environmental Biotechnology, School of Natural Sciences, University of Bangor, Wales, United Kingdom
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R. Mahadevan
1BioZone – Centre for Applied Bioscience and Bioengineering, Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON, Canada
4Institute of Biomedical Engineering, University of Toronto, Toronto, ON, Canada
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  • For correspondence: krishna.mahadevan@utoronto.ca
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ABSTRACT

Mine wastewater often contains dissolved metals at concentrations too low to be economically extracted by existing technologies, yet too high for environmental discharge. The most common treatment is chemical precipitation of the dissolved metals using limestone and subsequent disposal of the sludge in tailing impoundments. While it is a cost-effective solution to meet regulatory standards, it represents a lost opportunity. In this study, we engineered Escherichia coli to overexpress its native NikABCDE transporter and a heterologous metallothionein to capture nickel at concentrations in local effluent streams. We found the engineered strain had a 7-fold improvement in the bioaccumulation performance for nickel compared to controls, but also observed a drastic decrease in cell viability due to metabolic burden or inducer (IPTG) toxicity. Growth kinetic analysis revealed the IPTG concentrations used based on past studies lead to growth inhibition, thus delineating future avenues for optimization of the engineered strain and its growth conditions to perform in more complex environments.

Competing Interest Statement

The authors have declared no competing interest.

  • ABBREVIATIONS

    ABC
    ATP-binding cassette (transporter)
    SBP
    solute-binding protein
    IPTG
    isopropyl ß-D-1-thiogalactopyranoside
    NTP
    nucleotide triphosphate
    ICP-OES
    inductively coupled plasma optical emission spectrometry
    LB
    Luria-Bertani (nutrient media)
    kDa
    kilodaltons
    O.D.
    optical density
    NGR
    normalized growth ratio
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    Posted August 27, 2022.
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    Engineered nickel bioaccumulation in Escherichia coli by NikABCDE transporter and metallothionein overexpression
    P. Diep, H. Shen, J. A. Wiesner, N. Mykytczuk, V. Papangelakis, A. F. Yakunin, R. Mahadevan
    bioRxiv 2022.08.26.505507; doi: https://doi.org/10.1101/2022.08.26.505507
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    Engineered nickel bioaccumulation in Escherichia coli by NikABCDE transporter and metallothionein overexpression
    P. Diep, H. Shen, J. A. Wiesner, N. Mykytczuk, V. Papangelakis, A. F. Yakunin, R. Mahadevan
    bioRxiv 2022.08.26.505507; doi: https://doi.org/10.1101/2022.08.26.505507

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