Modification and Assembly of a Versatile Lactonase for Bacterial Quorum Quenching.

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TitleModification and Assembly of a Versatile Lactonase for Bacterial Quorum Quenching.
Publication TypeJournal Article
Year of Publication2018
AuthorsRhoads, MK, Hauk, P, Gupta, V, Bookstaver, ML, Stephens, K, Payne, GF, Bentley, WE
JournalMolecules
Volume23
Issue2
Date Published2018 Feb 06
ISSN1420-3049
Abstract

This work sets out to provide a self-assembled biopolymer capsule activated with a multi-functional enzyme for localized delivery. This enzyme,Pox, which is a lactonase and phosphotriesterase, provides a means of interrupting bacterial communication pathways that have been shown to mediate pathogenicity. Here we demonstrate the capability to express, purify and attachPox to the natural biopolymer chitosan, preserving its activity to "neutralize" long-chain autoinducer-1 (AI-1) communication molecules. Attachment is shown via non-specific binding and by engineering tyrosine and glutamine affinity 'tags' at the C-terminus for covalent linkage. Subsequent degradation of AI-1, in this case-(3-oxododecanoyl)-l-homoserine lactone (OdDHL), serves to "quench" bacterial quorum sensing (QS), silencing intraspecies communication. By attaching enzymes to pH-responsive chitosan that, in turn, can be assembled into various forms, we demonstrate device-based flexibility for enzyme delivery. Specifically, we have assembled quorum-quenching capsules consisting of an alginate inner core and an enzyme "decorated" chitosan shell that are shown to preclude bacterial QS crosstalk, minimizing QS mediated behaviors.

DOI10.3390/molecules23020341
Alternate JournalMolecules
PubMed ID29415497
Grant ListT32 AI089621 / AI / NIAID NIH HHS / United States