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Directed Protein Packaging within Outer Membrane Vesicles from Escherichia coli: Design, Production and Purification
Published on: November 16, 2016
Encapsulation of an enzyme cascade within the bacteriophage P22 virus-like particle
Dustin P Patterson1, Benjamin Schwarz, Ryan S Waters
1Department of Chemistry and Biochemistry, ‡Center for Bio-Inspired Nanomaterials, and §Deparment of Mathematical Sciences, Montana State University , Bozeman, Montana 59717, United States.
Abstract:
Developing methods for investigating coupled enzyme systems under conditions that mimic the cellular environment remains a significant challenge. Here we describe a biomimetic approach for constructing densely packed and confined multienzyme systems through the co-encapsulation of 2 and 3 enzymes within a virus-like particle (VLP) that perform a coupled cascade of reactions, creating a synthetic metabolon. Enzymes are efficiently encapsulated in vivo with known stoichiometries, and the kinetic parameters of the individual and coupled activities are characterized. From the results we develop and validate a mathematical model for predicting the expected kinetics for coupled reactions under co-localized conditions.
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