Part:BBa_K3596056
BIOT with RBS mut4 and Pilin gene mut4A
BIOT is a basic design for the expression of Geobacter sulfurreducens's pilin-based electrically conductive protein nanowires (e-PNs)[1]. The system includes a tac promoter which promotes the pilin gene(pilA) to express protein monomers; the monomers are later assembled into protein nanowires by the TypeIV pilin assembly system, which is constructed from the gene cluster at the downstream of the pilin gene(figure 1). The e-PNs can generate electric potential with the presence of water molecules in the air, which can be further modified for other purposes.
Figure 1: A Construction of BIOT gene circuit
Characterization
In order to improve the protein nanowire, we mutated the RBSs of the TypeIV pilin assembly system and the pilA gene to enhance the productivity and output voltage, respectively. This part shows the best combination effect, with the optimum RBS mutation RBS4 which increased the protein productivity by 34.1%(figure 2), and the optimum pilA mutation 4A which increased the electricity productivity by 46%(figure 3).
Figure 2: Different pilin productivity of engineered BIOT system with different RBS.
Figure 3: Different electricity productivity of engineered BIOT system with different pilin mutant.
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