Laminarihexaose

Twelve batches of Ganoderma lucidum (numbered S1 to S12) fruit bodies were collected from different places in China, including S1~S9 from Shouxiangu, Zhejiang Province, S10 from Huangshan, Anhui Province, and S11~S12 from Longquan, Zhejiang Province. The sample information is listed in Table 1. Trifluoroacetic acid (TFA), 1-phenyl-3-methyl-5-pyrazolone (PMP), monosaccharide standards (rhamnose, glucosamine, glucose, mannose, galactose, fucose, arabinose, fructose, xylose, glucuronic acid, and galacturonic acid), lipopolysaccharide (LPS), and polymyxin B were purchased from Sigma-Aldrich (St. Louis, MO, USA). Oligosaccharide standards including laminaribiose, laminaritriose, laminaritetraose, laminaripentaose, and laminarihexaose were purchased from Megazyme (Wicklow, Ireland). Pullulan standards P-5 (M_w = 6300 g/mol) and P-10 (M_w = 9800 g/mol) were purchased from Shodex (Tokyo, Japan). The RAW264.7 cell line was purchased from the Type Culture Collection of the Chinese Academy of Sciences. DMEM medium and fetal calf serum were bought from Gibco (Grand Island, NY, USA). The mouse TNF-α ELISA kit was bought from Beijing 4A Biotech Co., Ltd. (Beijing, China). All other reagents were analytical grade and produced in China.

M. loti EPS, S. meliloti EPS, and R. leguminosarum EPS ligands were obtained as described previously [22 (link),72 (link)]. Production and purification of the S. indica decasaccharide for binding assay was performed as described in [34 (link)]. L. digitata laminarin, maltodextrin, and chitin from shrimp shell were purchased from Sigma-Aldrich. Eisenia bicyclis laminarin, Pustulan, and Scleroglucan were purchased from Carbosynth. Laminarihexaose, laminaripentaose, and chitohexaose were purchased from Megazyme.