Nisin inducible production of listeriolysin O in Lactococcus lactis NZ9000

Simple item page
dc.contributor.authorBahey-El-Din, Mohammed
dc.contributor.authorGriffin, Brendan T.
dc.contributor.authorGahan, Cormac G.
dc.contributor.funderScience Foundation Irelanden
dc.date.accessioned2013-04-08T16:02:42Z
dc.date.available2013-04-08T16:02:42Z
dc.date.copyright2008
dc.date.issued2008-07
dc.description.abstractBackground: Listeria monocytogenes is a well-characterized food-borne pathogen that infects pregnant women and immunocompromised individuals. Listeriolysin O (LLO) is the major virulence factor of the pathogen and is often used as a diagnostic marker for detection of L. monocytogenes. In addition, LLO represents a potent antigen driving T cell-mediated immunity during infection. In the present work, Lactococcus lactis NZ9000 was used as an expression host to hyper-produce LLO under inducible conditions using the NICE (NIsin Controlled Expression) system. We created a modified pNZ8048 vector encoding a six-His-tagged LLO downstream of the strong inducible PnisA promoter. Results: The constructed vector (pNZPnisA:CYTO-LLO) was expressed in L. lactis NZ9000 and was best induced at mid-log phase with 0.2% v/v nisin for 4 h statically at 30°C. Purification of the His-tagged LLO was accomplished by Ni-NTA affinity chromatography and functionality was confirmed through haemolytic assays. Total LLO yield (measured as total protein content) was 4.43–5.9 mg per litre culture and the haemolytic activity was still detectable after 8 months of storage at 4°C. Conclusion: The LLO production method described in this work provides an approach to efficient LLO production in the Gram-positive Lactococcus bacterium to yield a significant source of the protein for research and diagnostic applications. Expression of LLO in L. lactis has a number of benefits over E. coli which may facilitate both in vivo and in vitro applications of this system.en
dc.description.sponsorshipScience Foundation Ireland (SFI-CSET);en
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationBahey-El-Din M, Griffin BT, Gahan CGM. (2008). Nisin inducible production of listeriolysin O in Lactococcus lactis NZ9000. Microbial Cell Factories, 7, 24.en
dc.identifier.doi10.1186/1475-2859-7-24
dc.identifier.issued24en
dc.identifier.journaltitleMicrobial Cell Factoriesen
dc.identifier.urihttps://hdl.handle.net/10468/1034
dc.identifier.volume7en
dc.language.isoenen
dc.publisherBioMed Centralen
dc.relation.urihttp://www.microbialcellfactories.com/content/7/1/24
dc.rights© 2008 Bahey-El-Din et al; licensee BioMed Central Ltd.en
dc.subjectListeria monocytogenesen
dc.subjectListeriolysin Oen
dc.subjectT cellen
dc.subjectLactococcus lactis NZ9000en
dc.subjectNICE (NIsin Controlled Expression) systemen
dc.subjectpNZ8048 vectoren
dc.subject.lcshMicrobiology.en
dc.titleNisin inducible production of listeriolysin O in Lactococcus lactis NZ9000en
dc.typeArticle (peer-reviewed)en
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
MBDB_nisinPV2008.pdf
Size:
514.83 KB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
license.txt
Size:
2.71 KB
Format:
Item-specific license agreed upon to submission
Description:
Download
Collections
APC Microbiome Ireland - Journal Articles
Microbiology - Journal Articles
Pharmacy - Journal Articles