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.
Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE, USA. wilfred@udel.edu.
Qing
Q
Sun
Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE, USA. wilfred@udel.edu.
Wilfred
W
Chen
D002482:Cellulose / Q000737:chemistry*; D044603:Cellulosomes / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D006868:Hydrolysis; D021141:Nucleic Acid Amplification Techniques
D002482:Cellulose; D004247:DNA