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Exopolysaccharides Produced by Lactobacillus rhamnosus KL 53A and Lactobacillus casei Fyos Affect Their Adhesion to Enterocytes

CORINNA KONIECZNA

CORINNA KONIECZNA

Department of Biotechnology and Food Microbiology, Poznan University of Life SciencesPoznan, Poland
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KONIECZNA, CORINNA
, 
MICHAŁ SŁODZIŃSKI

MICHAŁ SŁODZIŃSKI

Department of Biotechnology and Food Microbiology, Poznan University of Life SciencesPoznan, Poland
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SŁODZIŃSKI, MICHAŁ
 y 
MARCIN T. SCHMIDT

MARCIN T. SCHMIDT

Department of Biotechnology and Food Microbiology, Poznan University of Life SciencesPoznan, Poland
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SCHMIDT, MARCIN T.
  
04 sept 2018
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Categoría del artículo: original-paper
Publicado en línea: 04 sept 2018
Páginas: 273 - 281
Recibido: 01 feb 2017
Aceptado: 20 abr 2018
DOI: https://doi.org/10.21307/pjm-2018-032
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© 2018 Corinna Konieczna et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Adhesion efficiency of the fixed bacteria before degycosylation and after deglycosylation. LKL 53A, L. rhamnosus KL 53A, LcF, L. casei Fyos. The bars represent average percentages of bacterial cells that adhered to differentiated Caco-2 enterocytes, the whiskers represent average adhesion ± standard error.
Adhesion efficiency of the fixed bacteria before degycosylation and after deglycosylation. LKL 53A, L. rhamnosus KL 53A, LcF, L. casei Fyos. The bars represent average percentages of bacterial cells that adhered to differentiated Caco-2 enterocytes, the whiskers represent average adhesion ± standard error.

Fig. 2.

Normalized to 100% adhesion efficiency of L. casei Fyos grown in the presence of glycosidase inhibitors: castanospermine (Cas), deoxymannojirimycin hydrochloride (dMM), swainsonine (Swa), kifunensine (Kif), deoxynojirimycin hydrochloride (dNM) and without inhibitor (contr). The bars represent average percentages of bacterial cells that adhered to differentiated Caco-2 enterocytes, the whiskers represent average adhesion ± standard error. Statistically significant changes (p < 0.05) are marked with asterisks (*).
Normalized to 100% adhesion efficiency of L. casei Fyos grown in the presence of glycosidase inhibitors: castanospermine (Cas), deoxymannojirimycin hydrochloride (dMM), swainsonine (Swa), kifunensine (Kif), deoxynojirimycin hydrochloride (dNM) and without inhibitor (contr). The bars represent average percentages of bacterial cells that adhered to differentiated Caco-2 enterocytes, the whiskers represent average adhesion ± standard error. Statistically significant changes (p < 0.05) are marked with asterisks (*).

Fig. 3.

A comparison of relative protein number in the L. casei Fyos proteomic mass spectrometry data of the control cells (contr) and of the cells grown in the presence of swainsonine (Swa).
A comparison of relative protein number in the L. casei Fyos proteomic mass spectrometry data of the control cells (contr) and of the cells grown in the presence of swainsonine (Swa).

Fig. 4.

Changes in adhesion efficiency to Caco-2 cells of two lactic acid bacteria strains co-cultured with anaerobic gut bacteria. Adhesion efficiency was normalized to 100%. Statistically significant changes (p < 0.05) are marked with asterisks (*). C – control: either L. casei Fyos or L. rhamnosus KL 53A; LcF/FP, L. casei Fyos co-cultured with F. prausnitzii DSM 17677; LcF/BL, L. casei Fyos co-cultured with B. luti DSM 14534; LKL 53A/FP, L. rhamnosus KL 53A co-cultured with F. prausnitzii DSM 17677; LKL 53A/BL, L. rhamnosus KL 53A co-cultured with B. luti DSM 14534.
Changes in adhesion efficiency to Caco-2 cells of two lactic acid bacteria strains co-cultured with anaerobic gut bacteria. Adhesion efficiency was normalized to 100%. Statistically significant changes (p < 0.05) are marked with asterisks (*). C – control: either L. casei Fyos or L. rhamnosus KL 53A; LcF/FP, L. casei Fyos co-cultured with F. prausnitzii DSM 17677; LcF/BL, L. casei Fyos co-cultured with B. luti DSM 14534; LKL 53A/FP, L. rhamnosus KL 53A co-cultured with F. prausnitzii DSM 17677; LKL 53A/BL, L. rhamnosus KL 53A co-cultured with B. luti DSM 14534.

Composition of the Protein Deglycosylation Mix_

EnzymeHydrolyzed boundsNumber of enzymatic units added per 100 μl of reaction
O-GlycosidaseCatalyzes the removal of core 1 and core 3 O-linked disaccharides from glycoproteins.400000 U
PNGase FCleaves between the innermost GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides from N-linked glycoproteins unless α(1-3) core fucosylated  5000 U
NeuraminidaseCatalyzes the hydrolysis of terminal, non-reducing α2,3, α2,6, and α2,8 linked N-acetylneuraminic acid residues from glycoproteins and oligosaccharides.    500 U
β1-4 GalactosidaseCatalyzes the hydrolysis of terminal, non-reducing β1-4 linked D-galactopyranosyl residues from oligosaccharides and glycoproteins.      80 U
β-N-AcetylglucosaminidaseCatalyzes the hydrolysis of terminal, non-reducing β-N-Acetylglucosamine residues from oligosaccharides and glycoproteins.      40 U

Function of glycosidase inhibitors used to inhibit EPS synthesis in L_ casei Fyos_

GT inhibitorThe enzyme inhibited
Castanospermineβ-glucosidaseβ-xylosidaseα-mannosidaseβ-mannosidaseMaltase-glucoamylase
Deoxymannojirimycinα-(1→2)-mannosidase
Kifunensine
Swainsonineα-(1→3)-mannosidaseα-(1→6)-mannosidase
Deoxynojirimycin hydrochlorideα-glucosidaseMaltase-glucoamylase

Monosaccharides composition of the EPS produced by L_ rhamnosus KL 53A and L_ casei Fyos when grown on MRS agar (anaerobic conditions, 37°C, 18 h)_

Monomer% of a monomer in EPS isolated from the strain tested
Lactobacillus rhamnosus KL 53ALactobacillus casei Fyos
Maltose0.710
Glucose30.915.7
Galactose53.731.4
Arabinose14.651.6
Ribose01.4
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Ciencias de la vida, Microbiología y virología
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