Modification and Assembly of a Versatile Lactonase for Bacterial Quorum Quenching
This work sets out to provide a self-assembled biopolymer capsule activated with a multi-functional enzyme for localized delivery. This enzyme, SsoPox, which is a lactonase and phosphotriesterase, provides a means of interrupting bacterial communication pathways that have been shown to mediate patho...
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doaj-10500f0d75a141a5a1d5050209f01baa2020-11-24T23:57:54ZengMDPI AGMolecules1420-30492018-02-0123234110.3390/molecules23020341molecules23020341Modification and Assembly of a Versatile Lactonase for Bacterial Quorum QuenchingMelissa K. Rhoads0Pricila Hauk1Valerie Gupta2Michelle L. Bookstaver3Kristina Stephens4Gregory F. Payne5William E. Bentley6Institute for Bioscience and Biotechnology Research (IBBR), University of Maryland, College Park, MD 20742, USAInstitute for Bioscience and Biotechnology Research (IBBR), University of Maryland, College Park, MD 20742, USAFischell Department of Bioengineering, University of Maryland, College Park, MD 20742, USAFischell Department of Bioengineering, University of Maryland, College Park, MD 20742, USAInstitute for Bioscience and Biotechnology Research (IBBR), University of Maryland, College Park, MD 20742, USAInstitute for Bioscience and Biotechnology Research (IBBR), University of Maryland, College Park, MD 20742, USAInstitute for Bioscience and Biotechnology Research (IBBR), University of Maryland, College Park, MD 20742, USAThis work sets out to provide a self-assembled biopolymer capsule activated with a multi-functional enzyme for localized delivery. This enzyme, SsoPox, 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 attach SsoPox 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 N-(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.http://www.mdpi.com/1420-3049/23/2/341quorum quenchingantibioticsorganophosphateenzyme attachmentenzyme delivery |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Melissa K. Rhoads Pricila Hauk Valerie Gupta Michelle L. Bookstaver Kristina Stephens Gregory F. Payne William E. Bentley |
spellingShingle |
Melissa K. Rhoads Pricila Hauk Valerie Gupta Michelle L. Bookstaver Kristina Stephens Gregory F. Payne William E. Bentley Modification and Assembly of a Versatile Lactonase for Bacterial Quorum Quenching Molecules quorum quenching antibiotics organophosphate enzyme attachment enzyme delivery |
author_facet |
Melissa K. Rhoads Pricila Hauk Valerie Gupta Michelle L. Bookstaver Kristina Stephens Gregory F. Payne William E. Bentley |
author_sort |
Melissa K. Rhoads |
title |
Modification and Assembly of a Versatile Lactonase for Bacterial Quorum Quenching |
title_short |
Modification and Assembly of a Versatile Lactonase for Bacterial Quorum Quenching |
title_full |
Modification and Assembly of a Versatile Lactonase for Bacterial Quorum Quenching |
title_fullStr |
Modification and Assembly of a Versatile Lactonase for Bacterial Quorum Quenching |
title_full_unstemmed |
Modification and Assembly of a Versatile Lactonase for Bacterial Quorum Quenching |
title_sort |
modification and assembly of a versatile lactonase for bacterial quorum quenching |
publisher |
MDPI AG |
series |
Molecules |
issn |
1420-3049 |
publishDate |
2018-02-01 |
description |
This work sets out to provide a self-assembled biopolymer capsule activated with a multi-functional enzyme for localized delivery. This enzyme, SsoPox, 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 attach SsoPox 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 N-(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. |
topic |
quorum quenching antibiotics organophosphate enzyme attachment enzyme delivery |
url |
http://www.mdpi.com/1420-3049/23/2/341 |
work_keys_str_mv |
AT melissakrhoads modificationandassemblyofaversatilelactonaseforbacterialquorumquenching AT pricilahauk modificationandassemblyofaversatilelactonaseforbacterialquorumquenching AT valeriegupta modificationandassemblyofaversatilelactonaseforbacterialquorumquenching AT michellelbookstaver modificationandassemblyofaversatilelactonaseforbacterialquorumquenching AT kristinastephens modificationandassemblyofaversatilelactonaseforbacterialquorumquenching AT gregoryfpayne modificationandassemblyofaversatilelactonaseforbacterialquorumquenching AT williamebentley modificationandassemblyofaversatilelactonaseforbacterialquorumquenching |
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1725452894298177536 |