Lichenan from Icelandic moss, barley β-glucan, laminaribiose, laminaritriose, laminaritetraose, laminaripentaose, cellotriose, cellobiosyl-β-D-1,3-glucose (G4G3G) were purchased from Megazyme, Ireland. Gentiobiose and laminarin were purchased from Sigma Aldrich. All NMR analyses were performed at Panjab University, Chandigarh, India. All kinetic parameters were performed with GraphPad Prism.
Laminaritetraose
Manufactured by Megazyme
Sourced in Ireland
Laminaritetraose is a carbohydrate compound used as a standard in various analytical and research applications. It serves as a reference material for the identification and quantification of similar oligosaccharides.
Automatically generated - may contain errors
Lab products found in correlation
Laminaripentaose, by Megazyme (4 mentions)
Laminaritriose, by Megazyme (4 mentions)
Laminaribiose, by Megazyme (3 mentions)
Cellotriose, by Megazyme (2 mentions)
Gentiobiose, by Merck Group (1 mentions)
Laminarin, by Merck Group (1 mentions)
Barley β glucan, by Megazyme (1 mentions)
Laminarihexaose, by Megazyme (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Polymyxin b, by Merck Group (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Raw264.7 cell line, by National Collection of Authenticated Cell Cultures (1 mentions)
Laminaribiose, by Merck Group (1 mentions)
Ics 5000, by Thermo Fisher Scientific (1 mentions)
Costar spin x, by Corning (1 mentions)
Carbopac pa 100, by Thermo Fisher Scientific (1 mentions)
Glucose, by Merck Group (1 mentions)
Cellotetraose, by Megazyme (1 mentions)
Cellopentaose, by Megazyme (1 mentions)
6 protocols using laminaritetraose
1
Characterization of Carbohydrate Standards
2
Ganoderma lucidum Fruit Body Analysis
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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 (Mw = 6300 g/mol) and P-10 (Mw = 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.
3
Enzymatic Hydrolysis and HPAEC-PAD Analysis
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To exemplify the function of our enzymatic assay on an environmental sample, we hydrolyzed a sample stepwise and detected the resulting products via HPAEC with pulsed amperometric detection (PAD). The sample extract from Helgoland at April 27, 2017 was first hydrolyzed with 100 nM FbGH30 for 25 min at 37 °C, and then FaGH17A was applied in the same manner. Between each digestion, the reaction was stopped by boiling the sample for 5 min at 100 °C; precipitated protein was removed by filtration through 0.2-µm centrifuge filters (Costar Spin-X; Corning), and an aliquot was taken.
Samples were applied on an ICS-5000+ (Dionex) with electrochemical detection on a gold working electrode and a pH reference electrode (Ag/AgCl) according to Unfried et al. (37 (link)). Separation was attained by using a Dionex CarboPac PA100 analytical column at 35 °C. Glucose (Sigma), laminaribiose, laminaritriose, laminaritetraose, and laminaripentaose (all from Megazyme) were used as reference.
Samples were applied on an ICS-5000+ (Dionex) with electrochemical detection on a gold working electrode and a pH reference electrode (Ag/AgCl) according to Unfried et al. (37 (link)). Separation was attained by using a Dionex CarboPac PA100 analytical column at 35 °C. Glucose (Sigma), laminaribiose, laminaritriose, laminaritetraose, and laminaripentaose (all from Megazyme) were used as reference.
4
Preparation and Characterization of Oligosaccharides
5
Characterization of G. lucidum Supplements
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Nineteen batches (GL01 to GL19) of G. lucidum dietary supplements were purchased, in February or March 2015, directly from general e-commerce sites such as Amazon.com and eBay Inc. in the United States, and one batch of authenticated fruiting body of G. lucidum (GL20) was collected from Shandong Province of China (Table 1 ). Identity of G. lucidum fruiting body was confirmed by Professor Xiaolan Mao, Institute of Microbiology, China Academy of Sciences. The voucher specimens were deposited at the Institute of Chinese Medical Sciences, University of Macau, Macao, China.
D -glucose, α-amylase, starch (ST), and acetic anhydride were purchased from Sigma (St. Louis, MO, USA). Laminaribiose (DP2), laminaritriose (DP3), and laminaritetraose (DP4), β-1,3-D -glucan (GN), and β-1,3-D -glucanase were purchased from Megazyme (Wicklow, Ireland), and 8-aminonaphthalene-1,3,6-trisulphonic acid (ANTS) was purchased from Tokyo Chemical Industry (Tokyo, Japan). Silica gel 60 F254 TLC plates were obtained from Merck (Merck, Darmstadt, Germany). Polyacrylamide containing acrylamide/N, N-methylenebisacrylamide (19:1, w/w) was obtained from Bio-Rad (Hercules, CA, USA). Deionized water was prepared by a Millipore Milli-Q Plus system (Millipore, Bedford, MA, USA). All the other reagents were of analytical grade.
6
Alicyclobacillus sp. A4 Genomic Analysis
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The acidothermophilic Alicyclobacillus sp. A4 (whole genome sequenced) isolated from the hot spring water was stored in our laboratory [1 (link)]. GH3 β-glucosidase Bgl3A derived from Talaromyces leycettanus JCM12802 [376] was used as the reference. Ni2+-affinity beads from Suzhou Beaver Biomedical Engineering (China), and glucose oxidation (GOD-POD) kit from Beijing Leadman (China) were purchased. Substrates of p-nitrophenyl β-d -glucopyranoside (pNPG), p-nitrophenyl α-l -arabinofuranoside (pNPAf), p-nitrophenyl β-d -xylopyranoside (pNPX), barley β-glucan, lichenan, Avicel, and standard samples of daidzein, glycitein and genistein from Sigma-Aldrich (USA), cellobiose to cellohexose, laminaritetraose and maltose from Megazyme (Ireland), and daidzin from Tokyo Chemical Industry were obtained. The soybean meal was purchased from local market. All the other reagents were of analytical grade and commercially available.
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