Transformation efficiencies and transposition frequencies.

*<p>Transformation efficiencies and transposition frequencies were determined following electroporation of a replicative vector, namely pAM5, or a tetracycline-resistant TN5 transposome, respectively, into <i>B. breve</i> UCC2003 competent cells grown in different carbohydrates. Transposition events are average of independent duplicates.</p

Genomic position and surrounding regions of the insertion site of a number of transposon insertion mutants that were isolated based on their inability to grow on one or more carbohydrates.

<p>The diagram was drawn to scale using <i>B. breve</i> UCC2003 genome sequence information. White open arrowheads represent the location of the transposon with the specific location indicated as the genome coordinate based on accession number CP000303. Red arrows represent transposon-disrupted open reading frames. Grey arrows represent flanking open reading frames. Lollipops indicate transcriptional terminators predicted by ARNold Web server (<a href="http://rna.igmors.u-psud.fr/toolbox/arnold/" target="_blank">http://rna.igmors.u-psud.fr/toolbox/arnold/</a>).</p

Strains, plasmids and primers used.

<p>Restriction sites are underlined and a rho-transcriptional terminator sequence is highlighted in bold. NCFB, National Collection of Food Bacteria.</p

Growth profiles of <i>B.</i><i>breve</i> UCC2003 and derived transposon mutant strains, 101C6 (I), 181D10 (II) on lactose, galactose and ribose; and 164B7 (III) on lactose, pullulan and ribose.

<p>Presented data are average of duplicate independent growth experiments.</p

Confirmation of transposon insertion events into <i>B.</i><i>breve</i> UCC2003 and <i>B. breve</i> NCFB2258 genome by Southern hybridization.

<p>Blots of twenty-six randomly selected mutants of <i>B. breve</i> UCC2003 (Panel I, two blots) and sixteen randomly selected mutants of <i>B. breve</i> NCFB2258 (Panel II, single blot) are shown. Lanes labeled from A to R correspond to mutants whose insertion site was subsequently sequenced as indicated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064699#pone-0064699-t003" target="_blank">Table 3</a>.</p

MOESM1 of Cloning, expression and characterization of a β-d-xylosidase from Lactobacillus rossiae DSM 15814T

Additional file 1: Figure S1. Effect of pH (A) and temperature (B) on the β-xylosidase activity of Lactobacillus rossiae DSM 15814T; Table S1. Gene sequences BLAST alignment

<i>Bifidobacterium longum</i> general genome features.

<p><i>Bifidobacterium longum</i> general genome features.</p

B. longum 35624 EPS composition and structure.

<p>The structure is annotated as the chemical formula and in condensed form. Capital letters denote the residues as in Figs <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0162983#pone.0162983.g006" target="_blank">6</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0162983#pone.0162983.g007" target="_blank">7</a>.</p

Mild acid hydrolysis of EPS.

<p>(A) Separation of EPS fragments by PGC HPLC with MS/MS detection. The extracted ion chromatogram for mass 1008.39 Da shows four peaks. Their reducing end sugar was clearly revealed by ESI-MS/MS. Their assignment as either Gal or Glc and the interpretation in terms of fragment structures was done <i>a posteriori</i> based on MALDI-TOF data and on knowledge of the EPS structure. (B) Example of a MALDI-TOF/TOF fragment spectrum showing b-ions from the non-reducing and y- and y´ (= ^1,5x) -ions from the reducing end.</p

B. longum 35624 EPS characterization.

<p>(A) The 600 MHz ¹H NMR proton spectrum of the acid-treated <b>35624</b> EPS (D₂O, 338 K) is illustrated. A part of the high-field region is displayed in the insert. <b>(B)</b> Expansion plot of the 150 MHz 13C NMR spectrum of the acid-treated <b>35624</b> exopolysaccharide. The anomeric signals on the left confirmed the presence of a hexasaccharide repeat unit.</p