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Spontaneous Mutation Reveals Influence of Exopolysaccharide on Lactobacillus johnsonii Surface Characteristics

Tarih

2013

Yazar

Horn N.
Wegmann U.
Dertli E.
Mulholland F.
Collins S.R.A.
Waldron K.W.
Bongaerts R.J.
Mayer M.J.
Narbad A.

Üst veri

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Özet

As a competitive exclusion agent, Lactobacillus johnsonii FI9785 has been shown to prevent the colonization of selected pathogenic bacteria from the chicken gastrointestinal tract. During growth of the bacterium a rare but consistent emergence of an altered phenotype was noted, generating smooth colonies in contrast to the wild type rough form. A smooth colony variant was isolated and two-dimensional gel analysis of both strains revealed a protein spot with different migration properties in the two phenotypes. The spot in both gels was identified as a putative tyrosine kinase (EpsC), associated with a predicted exopolysaccharide gene cluster. Sequencing of the epsC gene from the smooth mutant revealed a single substitution (G to A) in the coding strand, resulting in the amino acid change D88N in the corresponding gene product. A native plasmid of L. johnsonii was engineered to produce a novel vector for constitutive expression and this was used to demonstrate that expression of the wild type epsC gene in the smooth mutant produced a reversion to the rough colony phenotype. Both the mutant and epsC complemented strains had increased levels of exopolysaccharides compared to the wild type strain, indicating that the rough phenotype is not solely associated with the quantity of exopolysaccharide. Another gene in the cluster, epsE, that encoded a putative undecaprenyl-phosphate galactosephosphotransferase, was deleted in order to investigate its role in exopolysaccharide biosynthesis. The ?epsE strain exhibited a large increase in cell aggregation and a reduction in exopolysaccharide content, while plasmid complementation of epsE restored the wild type phenotype. Flow cytometry showed that the wild type and derivative strains exhibited clear differences in their adhesive ability to HT29 monolayers in tissue culture, demonstrating an impact of EPS on surface properties and bacteria-host interactions. © 2013 Horn et al.

Kaynak

PLoS ONE

Cilt

8

Sayı

3

Bağlantı

https://dx.doi.org/10.1371/journal.pone.0059957
https://hdl.handle.net/20.500.12403/863

Koleksiyonlar

  • Scopus İndeksli Yayınlar Koleksiyonu [570]
  • WoS İndeksli Yayınlar Koleksiyonu [372]

İlgili Öğeler

Başlık, yazar, küratör ve konuya göre gösterilen ilgili öğeler.

  • Structure and biosynthesis of two exopolysaccharides produced by lactobacillus johnsonii FI9785 

    Dertli E.; Colquhoun I.J.; Gunning A.P.; Bongaerts R.J.; Le Gall G.; Bonev B.B.; Mayer M.J.; Narbad A. (2013)
    Exopolysaccharides were isolated and purified from Lactobacillus johnsonii FI9785, which has previously been shown to act as a competitive exclusion agent to control Clostridium perfringens in poultry. Structural analysis ...
  • Impact of exopolysaccharide production on functional properties of some Lactobacillus salivarius strains 

    Mercan E.; İspirli H.; Sert D.; Yılmaz M.T.; Dertli E. (Springer Verlag, 2015)
    The aim of this work was to characterize functional properties of Lactobacillus salivarius strains isolated from chicken feces. Detection of genes responsible for exopolysaccharide (EPS) production revealed that all strains ...
  • Characterization of a 4,6???glucanotransferase from Lactobacillus reuteri E81 and production of malto-oligosaccharides with immune-modulatory roles 

    İspirli H.; Şimşek Ö.; Skory C.; Sağdıç O.; Dertli E. (Elsevier B.V., 2019)
    A wide number of Lactic Acid Bacteria (LAB) species produce ?-glucans with their ability to synthesize glucansucrases (GS) which use sucrose as substrate for the glucan production. Recently another group of enzymes in LAB ...



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