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

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

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.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Outer Membrane Proteins / metabolism*
  • Cellulase / metabolism*
  • Cellulose / metabolism
  • Gram-Negative Bacteria / cytology
  • Gram-Negative Bacteria / metabolism
  • Hydrolysis
  • Nanoparticles / chemistry
  • Proteolipids / metabolism*

Substances

  • Bacterial Outer Membrane Proteins
  • Proteolipids
  • proteoliposomes
  • Cellulose
  • Cellulase

Grants and funding

The authors would like to acknowledge the funding support from National Science Foundation (CBET1263774). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.