Protecting enzymatic function through directed packaging into bacterial outer membrane vesicles

Bacteria possess innate machinery to transport extracellular cargo between cells as well as package virulence factors to infect host cells by secreting outer membrane vesicles (OMVs) that contain small molecules, proteins and genetic material. These robust proteoliposomes have evolved naturally to b...

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Bibliographic Details
Published in:Scientific reports 2016-04, Vol.6 (1), p.24866-24866, Article 24866
Main Authors: Alves, Nathan J., Turner, Kendrick B., Medintz, Igor L., Walper, Scott A.
Format: Article
Language:English
Subjects:
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Bacteria
Bacteria - enzymology
Bacteria - metabolism
Bacterial vesicles
Biodegradation
E coli
Environmental restoration
Enzyme Stability
Enzymes
Extracellular Vesicles - enzymology
Freeze Drying
Freeze-thawing
Genetic engineering
Gram-positive bacteria
Humanities and Social Sciences
Laboratories
Membrane vesicles
multidisciplinary
Nanoparticles
Packaging
Phosphoric Triester Hydrolases - metabolism
Phosphoric Triester Hydrolases - secretion
Phosphotriesterase
Plasmids
Proteins
Proteolipids - metabolism
Science
Storage conditions
Temperature
Time Factors
Virulence
Virulence factors
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Summary:Bacteria possess innate machinery to transport extracellular cargo between cells as well as package virulence factors to infect host cells by secreting outer membrane vesicles (OMVs) that contain small molecules, proteins and genetic material. These robust proteoliposomes have evolved naturally to be resistant to degradation and provide a supportive environment to extend the activity of encapsulated cargo. In this study, we sought to exploit bacterial OMV formation to package and maintain the activity of an enzyme, phosphotriesterase (PTE), under challenging storage conditions encountered for real world applications. Here we show that OMV packaged PTE maintains activity over free PTE when subjected to elevated temperatures (>100-fold more activity after 14 days at 37 °C), iterative freeze-thaw cycles (3.4-fold post four-cycles) and lyophilization (43-fold). We also demonstrate how lyophilized OMV packaged PTE can be utilized as a cell free reagent for long term environmental remediation of pesticide/chemical warfare contaminated areas.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep24866