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

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. * 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