Laminaribiose

The released oligosaccharides were separated by HPAEC on a CarboPacPA1 column (Dionex, Thermo Fisher Scientific, Waltham, MA, USA) equilibrated with 50 mm NaOAc in 0.2 m NaOH using a Dionex BioLC ICS 300 system (Dionex) equipped with a pulsed amperometric detector (PAD, Dionex, Thermo Fisher Scientific, Waltham, MA, USA). Oligosaccharides were eluted at 1 mL/min with a linear gradient of 50 mm NaOAc in 0.2 m NaOH to 350 mm NaOAc in 0.2 m NaOH over a 15 min period. Laminaribiose (Seikagaku, Tokyo, Japan) and cellodextrins (Sigma‐Aldrich, Merck, Darmstadt, Germany) were run as standards.

Restriction endonucleases, DNA ligase, and DNA polymerase were purchased from Takara Bio (Shiga, Japan). The QIAquick Kit was obtained from Qiagen (Hilden, Germany). 5-Bromo-4-chloro-3-indolyl-β-D-glucoside (X-glc) was purchased from Rose Scientific (Edmonton, AB, Canada). p-Nitrophenyl (pNP) α-D-galactopyranoside, pNP α-D-glucopyranoside, and pNP β-D-xylopyranoside were purchased from Nacalai (Kyoto, Japan). pNP α-D-mannopyranoside was purchased from Senn Chemicals (Zürich, Switzerland). The following chemicals were purchased from Sigma (St. Louis, MO, USA): avicel, pNP α-L-arabinofuranoside, pNP α-L-arabinopyranoside, pNP β-L-arabinopyranoside, pNP α-L-fucopyranoside, pNP β-D-fucopyranoside (pNPFuc), pNP β-D-galactopyranoside, pNP β-D-glucopyranoside (pNPGlc), pNP β-D-mannopyranoside, pNP N-acetyl-β-D-glucosaminide, pNP α-L-rhamnopyranoside, pNP α-D-xylopyranoside, and pNP β-D-cellobioside, sophorose, nigerose, maltose, isomaltose, lactose, and salicin. Cello-origosaccharides and laminaribiose were purchased from Seikagaku Kogyo (Tokyo, Japan). Gentiobiose was purchased from Tokyo Chemical Industry (Tokyo, Japan).