HaloTag mediated artificial cellulosome assembly on a rolling circle amplification DNA template for efficient cellulose hydrolysis

We report here the generation of four-component artificial cellulosomes onto a DNA scaffold using the self-labeling HaloTag for DNA conjugation. The resulting structures exhibited significantly improved cellulosome assembly as well as cellulose hydrolysis over compatible structures generated using protein scaffolds. Cellulose hydrolysis was further enhanced by 2-fold using the more complex cellulosome structures assembled onto DNA templates generated by rolling circle amplification (RCA). The flexibility to insert additional hybridization sites in a multiplexing manner using RCA should enable the assembly of a larger array of cellulases to better mimic the enzyme diversity of naturally occurring cellulosomes.