Synthetic Biology Engineering of Biofilms as Nanomaterials Factories.

Peter Nguyen1, Zsofia Botyanszki2, Tony Cho1, Pei-Kun Tay1, Peng Yin3 and Neel Joshi1, (1)Wyss Institute of Biologically Inspired Engineering, Harvard University, Cambridge, MA, (2)Department of Chemisty and Chemical Biology, Harvard University, Cambridge, MA, (3)Systems Biology, Harvard Medical School, Cambridge, MA

Programmable self-assembling living systems are the next generation of bioinspired advanced materials, and will integrate synthetic biology and materials science approaches towards reengineering abundant and robust biomaterials, such as bacterial biofilms. We have recently introduced â??Biofilm-Integrated Nanofiber Displayâ? (BIND) as a strategy for the programmable functionalization of the E. coli biofilm extracellular matrix by genetically fusing various peptide domains to the amyloidogenic protein CsgA, the key proteinaceous component of the biofilms. We find that these engineered CsgA fusion proteins are successfully secreted by the cellular export machinery and self-assemble into a network of extracellular amyloid nanofibers that displays the fusion peptide of interest in high density. The displayed peptide domains maintain their function and confer various non-natural functions to the biofilms as a whole. The BIND platform thus reconceptualizes the microbial communities as autonomous factories for the production of self-assembling bulk nanomaterials. Our results suggest that BIND is a novel strategy for the efficient broad functionalization of biofilms via engineered peptide or protein domains. Using this technology, large-scale structures could be constructed from on-demand nanomaterials with spatially programmable immobilized protein or peptides and controlled bulk mechanical properties.