B. infantis JCM 1222T (=ATCC 15697T) and B. kashiwanohense JCM 15439T were from Japan Collection of Microorganisms (RIKEN Bioresource Center, Tsukuba, Japan), while B. longum 105-A was a gift from Y. Kano at Kyoto Pharmaceutical University, Japan (currently available as JCM 31944 from RIKEN Bioresource Center). For clarity, only the locus tag numbers assigned to the ATCC 15697T (Blon_, GenBank accession no. CP001095.1) are used to describe the identical genes from B. infantis JCM 1222T. Bifidobacteria were anaerobically grown at 37°C in Gifu anaerobic medium (GAM; Nissui Pharmaceutical, Tokyo, Japan) or in de Man, Rogosa, and Sharpe medium (Becton Dickinson, Franklin Lakes, NJ, USA) containing 0.34% (w/v) sodium ascorbate and 0.02% (w/v) cysteine-HCl (MRS-CS) supplemented with 2% (w/v) Glc. When examining sugar utilization, MRS-CS broth was supplemented with 0.5% (w/v) of each sugar. Escherichia coli DH5α was used for genetic manipulations. Antibiotics and the concentrations used were as follows: chloramphenicol (Cm) (2.5 μg/ml for bifidobacteria and 10 μg/ml for E. coli), Sp (30 μg/ml for B. longum, 10 μg/ml for B. infantis, and 75 μg/ml for E. coli), and kanamycin (50 μg/ml for E. coli). InvivO2 400 (10% CO2, 10% H2, and 80% N2; Ruskinn Technology, Bridgend, UK) was used for anaerobic cultivation. Growth was monitored by measuring optical density at 600 nm (OD600) unless otherwise indicated.

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