Article Figures & Data
Figures
- FIG 1
Purified cranberry xyloglucan putative structures. Shown is the positive reflectron mode MALDI-TOF MS spectrum of cranberry xyloglucan obtained from a HiPrep Sephacryl S-100 HR size exclusion column. The voltage was set at 80 kV, and 500 profiles were collected. Major peaks at m/z 923, 953, 1,055, 1,085, 1,217, 1,247, and 1,379 represent sodium adducts ([M + Na]+) of xyloglucan with DPs ranging from 6 to 9 and sugar compositions as follows: [H3P3+ Na]+, m/z 923; [H4P2+ Na]+, m/z 953; [H3P4+ Na]+, m/z 1,055; [H4P3+ Na]+, m/z 1,085; [H4P4+ Na]+, m/z 1,217; [H5P3+ Na]+, m/z 1,247; [H5P4+ Na]+, m/z 1,379.
- FIG 2
Bacterial growth while utilizing cranberry xyloglucans. Growth curves of B. longum subsp. longum UCD401, L. plantarum ATCC BAA-793, L. johnsonii ATCC 33200, and B. longum subsp. infantis ATCC 15697 and JCM 1260 grown on mMRS medium containing 2% (wt/vol) xyloglucans. The curves are drawn from an average of three independent experiments.
- FIG 3
Bacterial fermentative end products of cranberry xyloglucan utilization. Shown are lactate (a), acetate (b), and formate (c) production; acetate/lactate ratios (d); and formate/lactate ratios (e). Bl, B. longum subsp. longum UCD401; Lp, Lactobacillus plantarum ATCC BAA-793. Averages of independent biological triplicates are shown, and bars represent the standard deviation of the mean. Organic acid production is expressed in millimolar absolute concentrations. Asterisks represent significant differences determined by two-way ANOVA and Sidak's multiple-comparison test (P < 0.05).
- FIG 4
Bacterial growth on additional fractions of cranberry xyloglucans. Growth curves represent B. longum subsp. infantis ATCC 15697, JCM 1260, JCM 1272, and JCM 7007; B. longum subsp. longum JCM 11347, ATCC 15708, and UCD401; Lactobacillus plantarum ATCC BAA-793; and Lactobacillusjohnsonii ATCC 33200 grown on mMRS medium containing 2% (wt/vol) xyloglucan fraction A2 (a) or 2% (wt/vol) xyloglucan fraction A6 (b). The curves are drawn from the average of at least three independent experiments, with the exception of strains JCM 1260, JCM 7007, JCM 11347, and JCM 15708 which are based on duplicates in panel a.
- FIG 5
Bifidobacterial fermentative end products of cranberry fraction A6 utilization. (a to c) Lactate (a), acetate (b), and formate (c) production while B. longum subsp. infantis JCM 1260 and B. longum subsp. longum UCD401 utilize xyloglucan fraction A6. (d) Acetate:lactate ratio after fermentation. Averages of independent biological triplicates are shown, and bars represent the standard deviation of the mean. Asterisks represent significant differences evaluated by two-way ANOVA and Sidak's multiple-comparison test (P < 0.05).
- FIG 6
Gene expression of B. longum subsp. longum UCD401 while utilizing cranberry fraction A6 as a sole carbon source. Genes in the arabinose cluster of B. longum subsp. longum UCD401 predicted to participate in xyloglucan metabolism are depicted on the x axis. Expression of four GH family genes (those for α-l-arabinofuranosidase, C-terminal [BL_0405], arabinan endo-1,5-α-l-arabinosidase [BL_0404, BL_0403], β-xylosidase [BL_0402]) (a) and those for a carbohydrate ABC transporter substrate-binding protein (BL_0398) and carbohydrate ABC transporter membrane proteins (BL_0397, BL_0396) (b) is shown as fold change relative to the control (glucose). Averages of independent biological triplicates are shown, and bars represent the standard deviation of the mean. Asterisks (* and **) represent significant differences compared to the control (glucose), evaluated by paired t test (P values of <0.05 and <0.005, respectively).
- FIG 7
Bacterial syntrophic interactions modeled with cranberry xyloglucans. Growth curves of B. longum subsp. infantis ATCC 15697 on mMRS medium containing xyloglucans (green) and supernatants from B. longum subsp. longum UCD401 (orange), L. plantarum ATCC BAA-793 (blue), and L. johnsonii ATCC 33200 (yellow) after xyloglucan fermentation. The curves are drawn from an average of three independent experiments.
Tables
- TABLE 1
Strains used in this study
Straina Species or subspecies Origin ATCC 15697 B. longum subsp. infantis Human infant feces JCM 1260 B. longum subsp. infantis Human infant feces JCM 1272 B. longum subsp. infantis Human infant feces JCM 7007 B. longum subsp. infantis Human infant feces JCM 11347 B. longum subsp. longum Human feces ATCC 15708 B. longum subsp. longum Human feces UCD401 B. longum subsp. longum Human feces ATCC BAA-793 L. plantarum Human saliva ATCC 33200 L. johnsonii Human blood ↵a UCD, University of California Davis Culture Collection; ATCC, American Type Culture Collection; JCM, Japanese Collection of Microorganisms.
- TABLE 2
Analysis of bacterial growth kinetics calculated with Wolfram Mathematica 10.3
Strain 2% xyloglucans 2% glucose k (h−1) ΔODasym tc (h) k (h−1) ΔODasym tc (h) B. infantis JCM 1260 NDa ND ND ND ND ND B. infantis ATCC 15697 ND ND ND ND ND ND B. longum UCD401 0.555 ± 0.055 0.15 ± 0.02b 19.7 ± 2.5d 0.614 ± 0.048 1.25 ± 0.09 13.7 ± 0.6 L. plantarum ATCC BAA-793 0.386 ± 0.049f 0.29 ± 0.03c 2.3 ± 0.60e 0.569 ± 0.005 1.48 ± 0.02 5.3 ± 0.60 L. johnsonii ATCC 33200 ND ND ND ND ND ND ↵a ND, not determined.
↵b Significant difference in the asymptotic OD value of B. longum UCD401 compared to L. plantarum and to the positive control, glucose (P < 0.05).
↵c Significant difference in the asymptotic OD value of L. plantarum compared to B. longum UCD401 and to the positive control, glucose (P < 0.05).
↵d Significant difference in the inflection point (tc) of B. longum UCD401 compared to L. plantarum and to the positive control (P < 0.05).
↵e Significant difference in the inflection point (tc) of L. plantarum compared to B. longum UCD401 (P < 0.05).
↵f Significant difference in the growth rate (k) of L. plantarum on xyloglucans compared to that of other strains on xyloglucans and glucose (P < 0.05).
- TABLE 3
Kinetic analysis of bacterial growth on A2 and A6 fraction xyloglucans calculated with Wolfram Mathematica 10.3
Strain Xyloglucans from fraction: A2 A6 k (h−1) ΔODasym tc (h) k (h−1) ΔODasyma tc (h) B. infantis JCM 1260 0.464 ± 0.280 0.17 ± 0.08 6.4 ± 3.3 1.188 ± 0.79 0.08 ± 0.02a 3.3 ± 1.8 B. infantis JCM 1272 0.427 ± 0.039 0.38 ± 0.10 12.4 ± 0.6 NDc ND ND B. infantis JCM 7007 0.215 ± 0.137 0.58 ± 0.34 17.6 ± 6.1 ND ND ND B. infantis ATCC 15697 NAd NA NA ND ND ND B. longum JCM 11347 ND ND ND ND ND ND B. longum ATCC 15708 ND ND ND ND ND ND B. longum UCD401 0.438 ± 0.152 0.37 ± 0.08 11.3 ± 3.0 0.270 ± 0.084 0.27 ± 0.03b 16.3 ± 0.6 L. plantarum ATCC BAA-793 NA NA NA 0.513 ± 0.061 0.25 ± 0.02b 2.9 ± 0.5 L. johnsonii ATCC 33200 NA NA NA 0.944 ± 0.166 0.12 ± 0.01a 1.4 ± 0.3 ↵a Significant difference in the asymptotic OD values for JCM 1260 and L. johnsonii compared to UCD401 and L. plantarum in A6 fraction treatment (P < 0.05).
↵b Significant difference in the asymptotic OD values for UCD401 and L. plantarum compared to JCM 1260 and L. johnsonii in A6 fraction treatment (P < 0.05).
↵c ND, not determined.
↵d NA, not available.
- TABLE 4
Growth kinetics of strains during syntrophic interaction
Strain Supernatant from: B. longum UCD401 L. plantarum ATCC BAA-793 L. johnsonii ATCC 33200 k (h−1) ΔODasym tc (h) k (h−1) ΔODasym tc (h) k (h−1) ΔODasym tc (h) B. infantis ATCC 15697 NDe ND ND 0.940 ± 0.106a 0.066 ± 0.008 10.3 ± 0.6 0.801 ± 0.077 0.093 ± 0.008 10.3 ± 0.6 B. longum UCD401 NAf NA NA 0.632 ± 0.120b 0.096 ± 0.005d 12.8 ± 0.6 0.468 ± 0.070d 0.162 ± 0.007c 11.0 ± 0.0 L. plantarum ATCC BAA-793 0.890 ± 0.158d 0.115 ± 0.009d 2.1 ± 0.9 NA NA NA NA NA NA ↵a Significant difference between the growth rates of B. infantis ATCC 15697 in different conditioned media determined by paired t test (P < 0.05).
↵b Significant difference between the growth rates of B. longum UCD401 in different conditioned media determined by paired t test (P < 0.05).
↵c Significant difference between the asymptotic OD values of B. longum UCD401 in different conditioned media determined by paired t test (P < 0.05).
↵d Significant difference in the growth kinetics of each strain in different conditioned media and its growth kinetics in xyloglucans alone (shown in Table 2) determined by paired t test (P < 0.05).
↵e ND, not determined.
↵f NA, not available.
- TABLE 5
Primers used in this study
Supplemental material
- Supplemental file 1 -
Elution profile of cranberry xyloglucans on a Dephacryl S-100 HR 16/60 column eluted with deionized water (Fig. S1); 1H NMR spectra of cranberry xyloglucan (Fig. S2); LC-MS analysis of xyloglucans before fermentation and remaining post-fermentation by B. longum subsp. longum UCD 401 and Lactobacillus plantarum ATCC BAA-793 (Fig. S3); FL-HPLC chromatograms of xyloglucans before fermentation (control) and remaining post-fermentation by B. longum subsp. longum UCD 401 (Fig. S4); LC-MS analysis for specific xyloglucan structures by B. longum subsp. longum UCD 401 fermentation (Fig. S5); xyloglucan glycoprofile of B. longum subsp. longum UCD 401 grown on a medium supplemented with 2% (wt/vol) cranberry xyloglucans (Fig. S6); comparison of utilizations of different xyloglucan fractions (Fig. S7); LC-MS analysis of xyloglucans remaining after syntropic interaction of B. longum subsp. infantis ATCC 15697 with Lactobacillus plantarum ATCC BAA-793, Lactobacillus plantarum ATCC BAA-793, or B. longum subsp. longum UCD 401 (Fig. S8); genomic regions putatively linked with xyloglucan/arabinoxylose utilization in B. longum strains (Fig. S9); growth curves of B. longum subsp. infantis JCM 1260, B. longum subsp. infantis JCM 1272, B. longum subsp. infantis JCM 7007, B. longum subsp. longum JCM 11347, B. longum subsp. longum ATCC 15708, and B. longum subsp. longum UCD 401 on modified MRS containing 2% (wt/vol) arabinose (Fig. S10); MALDI-TOF mass spectrometry of cranberry xyloglucans with sodium adduct ions: composition and tentative assignments (Table S1).
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- Supplemental file 1 -