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PLoS One. 2014 May 12;9(5):e97103. doi: 10.1371/journal.pone.0097103. eCollection 2014.

Positional assembly of enzymes on bacterial outer membrane vesicles for cascade reactions.

Author information

1
Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, United States of America.
2
School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York, United States of America.

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.

PMID:
24820175
PMCID:
PMC4018249
DOI:
10.1371/journal.pone.0097103
[Indexed for MEDLINE]
Free PMC Article

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