title: Positional assembly of enzymes on bacterial outer membrane vesicles for cascade reactions.
abstract: The systematic organization of enzymes is a key feature for the efficient operation of cascade reactions in nature. Here, we demonstrate a facile method to create nanoscale enzyme cascades by using engineered bacterial outer membrane vesicles (OMVs) that are spheroid nanoparticles (roughly 50 nm in diameter) produced by Gram-negative bacteria during all phases of growth. By taking advantage of the fact that OMVs naturally contain proteins found in the outer cell membrane, we displayed a trivalent protein scaffold containing three divergent cohesin domains for the position-specific presentation of a three-enzyme cascade on OMVs through a truncated ice nucleation protein anchoring motif (INP). The positional assembly of three enzymes for cellulose hydrolysis was demonstrated. The enzyme-decorated OMVs provided synergistic cellulose hydrolysis resulting in 23-fold enhancement in glucose production than free enzymes.
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, United States of America.
authors.forename: Miso
authors.initials: M
authors.lastname: Park
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, United States of America.
authors.forename: Qing
authors.initials: Q
authors.lastname: Sun
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, United States of America.
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
authors.affiliation: School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York, United States of America.
authors.forename: Matthew P
authors.initials: MP
authors.lastname: DeLisa
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, United States of America.
authors.forename: Wilfred
authors.initials: W
authors.lastname: Chen
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D002480:Cellulase / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism; D006090:Gram-Negative Bacteria / Q000166:cytology / Q000378:metabolism; D006868:Hydrolysis; D053758:Nanoparticles / Q000737:chemistry; D011510:Proteolipids / Q000378:metabolism*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D011510:Proteolipids; C020485:proteoliposomes; D002482:Cellulose; D002480:Cellulase