D xylose

The methods of pentose (D(+)-Xylose, Wako, Osaka, Japan) and hexose (D-(+)-Glucose, Sigma-Aldrich, St. Louis, MO, USA) measurement were modified based on the phenol-sulfuric acid method of Nielsen [23 ]. Briefly, the sample was extracted by deionized water at 95 °C for 30 min and cooled down to room temperature before use. After that, 1 mL 5% phenol solution and 5 mL sulfuric acid were added to PWMC, FPW, and PP water extracts. After incubation for 15 min, the absorbance was measured at 480 nm with U-2900 Spectrophotometer (Hitachi, Tokyo, Japan) for the standard curveof xylose and glucose

d-Allose, d-glucose, α-Glc1P, d-gluconic acid, lactose, d-mannose, and d-xylose were purchased from Fujifilm Wako Pure Chemical (Osaka, Japan); N-acetyl-d-glucosamine, d-galactose, and sucrose were purchased from Nacalai Tesque (Kyoto, Japan); d-glucosamine was purchased from Tokyo Chemical Industry (Tokyo, Japan); cellobiose, d-galacturonic acid, and d-glucuronic acid were purchased from Sigma (St. Louis, MO, USA). β-(1 → 4)-Mannobiose was prepared as previously described^{29 (link)}. Lactoless L3 (β-galactosidase) was provided by Daiwa Kasei (Shiga, Japan). SP from Bifidobacterium longum was prepared according to the method described by Nishimoto and Kitaoka^{5 (link)}. One unit of SP was defined as the amount of enzyme required to phosphorolyze 1 μmol of sucrose in 1 min. SP activity was measured as follows: a reaction mixture (50 μL) containing appropriate concentration of enzyme, 20 mM sucrose, 40 mM sodium phosphate, 100 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES)-NaOH buffer (pH 7.0), and 0.2 mg/mL bovine serum albumin (BSA, Nacalai Tesque) was incubated at 37 °C for 10 min. The enzymatic reaction was terminated by incubating the sample at 80 °C for 3 min, and the liberated d-fructose was measured using a d-Fructose/d-Glucose Assay Kit (Megazyme, Brey, Ireland).