Peptide-mediated microalgae harvesting method for efficient biofuel production

Background: Production of biofuels from microalgae has been recognized to be a promising route for a sustainable energy supply. However, the microalgae harvesting process is a bottleneck for industrialization because it is energy intensive. Thus, by displaying interactive protein factors on the cell wall, oleaginous microalgae can acquire the autoand controllable-flocculation function, yielding smarter and energy-efficient harvesting. Results: Towards this goal, we established a cell-surface display system using the oleaginous diatom Fistulifera solaris JPCC DA0580. Putative cell wall proteins, termed frustulins, were identified from the genome information using a homology search. A selected frustulin was subsequently fused with green fluorescent protein (GFP) and a diatom cell-surface display was successfully demonstrated. The antibody-binding assay further confirmed that the displayed GFP could interact with the antibody at the outermost surface of the cells. Moreover, a cell harvesting experiment was carried out using silica-affinity peptide-displaying diatom cells and silica particles where engineered cells attached to the silica particles resulting in immediate sedimentation. Conclusion: This is the first report to demonstrate the engineered peptide-mediated harvesting of oleaginous microalgae using a cell-surface display system. Flocculation efficiency based on the silica-affinity peptide-mediated cell harvesting method demonstrated a comparable performance to other flocculation strategies which use either harsh pH conditions or expensive chemical/biological flocculation agents. We propose that our peptide-mediated cell harvest method will be useful for the efficient biofuel production in the future.