Adhesion of the genome-sequenced Lactococcus lactis subsp cremoris IBB477 strain is mediated by specific molecular determinants
Radziwill-Bienkowska, Joanna Maria | Le, Doan Thanh Lam | Szczesny, Pawel | Duviau, Marie-Pierre | Aleksandrzak-Piekarczyk, Tamara | Loubière, Pascal | Mercier-Bonin, Muriel | Bardowski, Jacek Karol | Kowalczyk, Magdalena | Institute of Biochemistry and Biophysics ; Pawinskiego 5a | Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP) ; Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse) ; Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS) | Faculty of Biology ; Uniwersytet im. Adama Mickiewicza w Poznaniu = Adam Mickiewicz University in Poznań (UAM) | The 'Studies of nucleic acids and proteins-from basic to applied research' project is realised within the International PhD Projects Programme of the Foundation for Polish Science (MPD/2009-3/2). The project is cofinanced by the EU-Regional Development Fund. This work was also funded by the European Funds Portal Innovative Economy 'Centre of medicinal product biotechnology. Package of innovative biopharmaceuticals for human and animal therapy and prophylactics' WND-POIG.01.01.02-00-007/08
Understanding the nature of mucus-microbe interactions will provide important information that can help to elucidate the mechanisms underlying probiotic adhesion. This study focused on the adhesive properties of the Lactococcus lactis subsp. cremoris IBB477 strain, previously shown to persist in the gastrointestinal tract of germ-free rats. The shear flow-induced detachment of L. lactis cells was investigated under laminar flow conditions. Such a dynamic approach demonstrated increased adhesion to bare and mucin-coated polystyrene for IBB477, compared to that observed for the MG1820 control strain. To identify potential genetic determinants giving adhesive properties to IBB477, the improved high-quality draft genome sequence comprising chromosome and five plasmids was obtained and analysed. The number of putative adhesion proteins was determined on the basis of surface/extracellular localisation and/or the presence of adhesion domains. To identify proteins essential for the IBB477 specific adhesion property, nine deletion mutants in chromosomal genes have been constructed and analysed using adhesion tests on bare polystyrene as well as mucin-, fibronectin- or collagen IV-coated polystyrene plates in comparison to the wild-type strain. These experiments demonstrated that gene AJ89_07570 encoding a protein containing DUF285, MucBP and four Big_3 domains is involved in adhesion to bare and mucin-coated polystyrene. To summarise, in the present work, we characterised the adhesion of IBB477 under laminar flow conditions; identified the putative adherence factors present in IBB477, which is the first L. lactis strain exhibiting adhesive and mucoadhesive properties to be sequenced and demonstrated that one of the proteins containing adhesion domains contributes to adhesion.
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