Figures & data
Notes: (a, b) The growth curve of BO and BC grown on 2.5 and 5 mg/mL cellobiose as the sole carbon source, respectively, was tested. (c) The growth curve of probiotics grown on 5 mg/mL cellobiose were detected. (d, e) The growth curve of BO and BC grown on 2.5 and 5 mg/mL cellotetrose as the sole carbon source, respectively, were analyzed. (f) Probiotics grew on 5 mg/mL cellotetrose were measured. Data represent technical triplicates (n = 3). Abbreviation: BO (Bacteroides ovatus), BC (Bacteroides cellulosilyticus), LR (Lactobacillus reuteri), LGG (Lactobacillus rhamnosus) and BL (Bifidobacterium longum).
(a) The supernatants of BO grown on cellobiose were analyzed. (b) BO was cultured either with glucose or cellobiose, and cells were collected till log phase, respectively. Then, cells were incubated with cellobiose, and the end products were analyzed. (c) Cells were treated with or without proteinase K for 6 h and then incubated with cellobiose for analyzing. (d) Collected cells were treated with or without proteinase K for 12 h and then incubated with cellobiose for analyzing. 1 means glucose; 2 means cellobiose.
Notes: (a) Up-regulated genes were selected based on Log2FC ≥ 3 and showed by heatmap. (b, c) The up-regulated genes were arranged into two PULs and confirmed by qPCR. (d) Schematic diagram of PUL1. (e) Schematic diagram of PUL2. Data represent technical triplicates (n = 3).
Notes: (a) Cellobiose activated enzymes and glycan binding proteins were tested by Western blotting. (b, c) Proteins cellar location was measured by immunofluorescence and Western blotting. PK means proteinase K. (d – f) Enzymatic activity of BACOVA_02626GH5, BACOVA_026-30GH5 and BACOVA_02745GH3 were analyzed by HPAEC-PAD.
Notes: (a) The structure of BACOVA_02626GH5 was predicted by AlphaFold2. (b) The structure of BACOVA_02630GH5 was predicted by AlphaFold2. (c, d) The catalytic pocket of BACOVA_02626GH5 was predicted by point-site. The catalytic residues of BACOVA_02626 was Glu 154 and Glu 241 (the orange arrow showed). (e, f) The catalytic pocket of BACOVA_02630GH5 was predicted by point-site. The catalytic residues of BACOVA_02630 was Glu 181 and Glu 293 (the blue arrow showed). Binding predication of glucose with cellulase (g) BACOVA_02626GH5 and (h) BACOVA_02630GH5 by docking.
(a) Schematic diagram of animal test. (b) Body weight of mice. (c) sobs index. (d, e) Principal component analysis (PCA) plots of fecal microbiota based on the ANOSIM and Adonis, respectively, were analyzed on genus level (R = 0.2153, p = 0.009). (f) The composition of gut microbiota was analyzed based on the top 50 species at genus level showed by Heatmap. (g) LEfSe analysis on genus level was analyzed (LDA = 2.5). (h) Compared cellobiose group with vehicle group on genus level. Data were presented as the mean ± SD, and n = 10.
(a) Functional activity of bacteria was predicted by PICRUSt2 tool. (b) Carbohydrate related enzymes were predicted by Tax4Fun tool. (c) KEGG pathways were predicted by PICRUSt1 tool. (d – f) mRNA expression level of TNF-α, IL-6 and H2R from colonic tissue were measured by qPCR. (g) Protein level of colonic TNF-α and IL-6 were analyzed by IHC. Data represent technical triplicates (n = 10).
Supplemental material