Construction of Cupriavidus necator displayed with superoxide dismutases for enhanced growth in bioelectrochemical systems

Abstract

It is of great significance to utilize CO₂ as feedstock to synthesize biobased products, particularly single cell protein (SCP) as the alternative food and feed. Bioelectrochemical system (BES) driven by clean electric energy has been regarded as a promising way for Cupriavidus necator to produce SCP from CO₂ directly. At present, the key problem of culturing C. necator in BES is that reactive oxygen species (ROS) generated in cathode chamber are harmful to bacterial growth. Therefore, it is necessary to find a solution to mitigate the negative effect of ROS. In this study, we constructed a number of C. necator strains displayed with superoxide dismutase (SOD), which allowed the decomposition of superoxide anion radical. The effects of promoters and signal peptides on the cell surface displayed SOD were analyzed. The proteins displayed on the surface were further verified by the fluorescence experiment. Finally, the growth of C. necator CMS incorporating a pBAD-SOD-E-tag-IgAβ plasmid could achieve 4.9 ± 1.0 of OD₆₀₀ by 7 days, equivalent to 1.7 ± 0.3 g/L dry cell weight (DCW), and the production rate was 0.24 ± 0.04 g/L/d DCW, around 2.7-fold increase than the original C. necator CMS (1.8 ± 0.3 of OD₆₀₀). This study can provide an effective and novel strategy of cultivating strains for the production of CO₂-derived SCP or other chemicals in BES.

Keywords

Carbon dioxide / Single cell protein / Bioelectrochemical system / Cell surface display / Hydrogen-oxidizing bacteria

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Funding

Key Technologies Research and Development Program(2021YFD1301002)

Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project(TSBICIPKJGG-012-04)