Fucose

Manufactured by Merck Group

Sourced in United States, Germany, China, Sao Tome and Principe

Fucose is a monosaccharide commonly found in glycoproteins and glycolipids. It serves as a core component in various biological processes.

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93 protocols using fucose

Fucose Quantification in Murine Brains

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Mice of ages indicated were anesthetized and perfused through the heart with PBS, brain tissues isolated and frozen at −80°C until used. Whole brains were homogenized in water, sonicated and an aliquot saved for protein concentration. The remaining lysate was adjusted to pH 1.0 with 1M H₂SO₄ (Sigma, St. Louis, MO), heated in a sand bath at 100°C for two h, neutralized with barium hydroxide (Sigma, St. Louis, MO), and the volume adjusted to 1 mL with water. After 30 min at 4°C, samples were spun down at 25,000Xg for 10 min and the supernatant isolated and stored at −20°C. Fucose content was determined by the Dische assay [18] (link) with known concentrations of Fucose (Sigma, St. Louis, MO) used as a standard. Each sample was normalized to protein concentration and the data plotted as nmol Fucose/mg total protein.

Idol R.A., Wozniak D.F., Fujiwara H., Yuede C.M., Ory D.S., Kornfeld S, & Vogel P. (2014). Neurologic Abnormalities in Mouse Models of the Lysosomal Storage Disorders Mucolipidosis II and Mucolipidosis III γ. PLoS ONE, 9(10), e109768.

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Fecal Fucose Degradation Assay

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To evaluate fucose degradation activity, stool samples were cultured as previously reported with some modifications^{75 (link)}. Briefly, fecal samples stored at –80 °C were thawed on ice and resuspended in phosphate buffer (0.1 M, pH 7.0) at 32% (wt/vol) to prepare the inoculum. Fermentation medium (14 mg/mL peptone, 0.3 mg/mL cysteine, 0.3 mg/mL sodium sulfide, and 1.2 μg/mL Resazurin) was supplemented with or without 50 mg/mL fucose (Sigma Aldrich), and was deoxygenated in a jar containing AnaeroPack as an oxygen absorber (A-03, Mitsubishi Gas Chemical), for 24 h before use. The inoculum (50 μL) and the fermentation medium (180 μL) were mixed in a 14 mL tube and incubated at 37 °C with shaking under anaerobic conditions in a jar containing AnaeroPack for 20 h. The reaction was stopped by incubating the culture tube on ice for 1 h, and fucose concentration was measured using L-fucose Kit (Megazyme) in accordance with the manufacturer’s instructions.

Hirota M., Tamai M., Yukawa S., Taira N., Matthews M.M., Toma T., Seto Y., Yoshida M., Toguchi S., Miyagi M., Mori T., Tomori H., Tamai O., Kina M., Sakihara E., Yamashiro C., Miyagi M., Tamaki K., Wolf M., Collins M.K., Kitano H, & Ishikawa H. (2023). Human immune and gut microbial parameters associated with inter-individual variations in COVID-19 mRNA vaccine-induced immunity. Communications Biology, 6, 368.

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Extraction and Analysis of Himalayan Plant Polysaccharides

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Roots of Mirabilis himalaica (Edgew) heim were purchased from Qinghai Tibetan hospital in Qinghai (batch number: 20161023; Xining, Qinghai province, China). Dextran standards (5800, 11 800, 47 300, 100 000, 380 000, and 788 000 Da) were acquired from Pharmacia Biotech (Uppsala, Sweden). Standard monosaccharides (glucose, d-glucosamine hydrochloride, mannose, rhamnose, ribose, galactose, fucose, N-acetyl-d-glucosamine, d-galactosamine hydrochloride, xylose, fructose, arabinose, guluronic acid, mannuronic acid, galacturonic acid, glucuronic acid) were purchased from Sigma chemical. Dialysis membranes (size 36, MW_CO = 8–14 kDa) were purchased from Wako. Acetic acid, acetic anhydride, sodium borohydride, chloroform, NaOH powder, DMSO, iodomethane, formic acid, methanol, trifluoroacetic acid were all analytical reagents.

Bo S., Dan M., Han W., Ochir S., Bao L., Liu L., Muschin T, & Baigude H. (2022). Physicochemical properties, immunostimulatory and antioxidant activities of a novel polysaccharide isolated from Mirabilis himalaica (Edgew) Heim. RSC Advances, 12(27), 17264-17275.

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Characterization of Helvella leucopus

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The mature fruiting body samples were purchased in Bachu county, Xinjiang province, China and were identified by Guangdong Institute of Microbiology to be Helvella leucopus. Petroleum ether, anhydrous ethanol, and trichloromethane were procured from Sinopharm Chemical Reagent (Chemical Reagent Co., ltd., Beijing, China). The standards fucose, rhamnose, arabinose, galactose, glucose, xylose, mannose, fructose, and ribose were acquired from Sigma-Aldrich (St. Louis, MO, USA). In addition, RNA-easy Isolation Reagent, HiScript III-RT SuperMix, and ChamQ Universal SYBR qPCR Master Mix (Vazyme Biotech Co. ltd. Nanjing, Jiangsu, China) were used in this study.

Ge Y., Qiu H, & Zheng J. (2022). Physicochemical characteristics and anti-hyperlipidemic effect of polysaccharide from BaChu mushroom (Helvella leucopus). Food Chemistry: X, 15, 100443.

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Formulation of Basal Cell Culture Media

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For basal media, Dulbecco's modified Eagle's medium (DMEM) with HEPES, and Coon's modified Ham's F-12 with HEPES (F12) were obtained from Sigma-Aldrich. DMEM without calcium chloride was obtained from Life Technologies (Carlsbad, CA). KBM-Gold™ (keratinocyte basal medium, calcium ion-free) from Lonza (Walkersville, MD).
Unless otherwise stated, the following reagents were used as purchased from Sigma-Aldrich: Alanyl-glutamine, bovine serum albumin, calcium chloride, cholesterol, ethanolamine, fructose, fucose, ga lactose, glucose, glucosamine, gluconic acid, glucuronic acid, glutathione (reduced), heparin, hydrocortisone, hypoxanthine, lactose, LONG®R³ IGF-1, manganese (II) chloride, non-essential amino acids, oxalacetic acid, progesterone (soluble), putrescine, retinyl acetate (soluble), sodium pyruvate, sodium selenite, TAPSO (N-[Tris(hydroxymethyl)methyl]-3-amino-2-hydroxypropanesulfonic acid), taurine, thioglycerol, partially saturated human transferrin, triiodothyronine (T3), uridine, and vitamin E acetate. Other reagents and sources were as follows: Ascorbic acid phosphate, Mg salt (Wako Chemicals, Richmond, VA); ammonium molybdate (Fisher Scientific, Suwanee, GA); bovine calf serum (CS; Lonza); carnitine tartrate (LKT Laboratories, St. Paul, MN); linoleic acid (Cayman Chemical, Ann Arbor, MI); thiamine and thymidine (Santa Cruz Biotechnology, Dallas, TX).

Pfeffer B.A., Xu L., Porter N.A., Rao S.R, & Fliesler S.J. (2016). Differential Cytotoxic Effects of 7-Dehydrocholesterol-derived Oxysterols on Cultured Retina-derived Cells: Dependence on Sterol Structure, Cell Type, and Density. Experimental eye research, 145, 297-316.

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Auricularia polytricha Cultivation Protocol

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Auricularia polytricha No.10 strain was purchased from Jiangsu Tianda Institute of Edible Fungi (Jiangsu, China).
Bovine serum albumin (BSA), Coomassie brilliant blue G–250, and ascorbic acid were obtained from Aladdin Reagent Co., Ltd. (Shanghai, China). Concentrated sulfuric acid, phosphoric acid, salicylic acid, trichloroacetic acid (TFA), anhydrous alcohol, acetone, anhydrous ether, glucose, sucrose, fructose, maltose and lactose, urea, peptone, ammonium sulfate, yeast and corn starch, neutral protease, 30% hydrogen peroxide (H₂O₂), phenol, Vitamin C (Vc) sodium hydroxide, ferrous sulfate (FeSO₄), ferric chloride (FeCl₃), sodium dihydrogen phosphate, disodium hydrogen phosphate, and potassium ferricyanide [K₃Fe(CN)₆] were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). The monosaccharide Standards including fucose (Fuc), rhamnose (Rha), arabinose (Ara), mannose (Man), galactose (Gal), glucose (Glc), xylose (Xyl), fructose (Fru), galacturonic acid (GalA), glucuronic acid (GlcA), 2,2–diphenyl–1–picrylhydrazyl (DPPH), and potassium bromide (KBr, Spectrum pure grade) were purchased from Sigma Chemical Co. (St. Louis, MO, United States). All chemicals and solvents used in the current work were of analytical grade.

Yang Z., Hu Y., Wu J., Liu J., Zhang F., Ao H., Zhu Y., He L., Zhang W, & Zeng X. (2022). High-Efficiency Production of Auricularia polytricha Polysaccharides Through Yellow Slurry Water Fermentation and Its Structure and Antioxidant Properties. Frontiers in Microbiology, 13, 811275.

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Quantification of Human Milk Oligosaccharides

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fucose, 2′-FL, and 3-FL were quantified from the simulation units to which HMOs were added as per Salli et al. [26 (link)] with modifications. Standard solutions of fucose (Sigma-Aldrich, St. Louis, MO, USA), 2′-FL, and 3-FL were prepared in water to concentrations of 80, 60, 40, 20, and 10 mg/l and stored at +4 °C. Sample solutions were centrifuged at 16,000× g for 5 min; then, 50 μL of the supernatant and 200 μL of ethanol were mixed in a microcentrifuge tube and incubated at +4 °C for 30 min. After centrifugation at 16,000× g for 5 min, 200 μL of the supernatant was evaporated to dryness at 30 °C under stream of nitrogen, and the solid residue was dissolved in 2000 μL of water and filtered. Separation and detection of the analytes were performed using high-performance anion-exchange chromatography as previously described [26 (link)]. Retention times of fucose, 2′-FL, and 3-FL were 6.3 min, 26 min, and 18 min, respectively.

Salli K., Hirvonen J., Anglenius H., Hibberd A.A., Ahonen I., Saarinen M.T., Maukonen J, & Ouwehand A.C. (2023). The Effect of Human Milk Oligosaccharides and Bifidobacterium longum subspecies infantis Bi-26 on Simulated Infant Gut Microbiome and Metabolites. Microorganisms, 11(6), 1553.

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Biochemical Assay Reagent Preparation

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Glycine, glucose, galactose, arabinose, fucose, mannose, rhamnose, glucuronic acid and xylose gallic acid, 3,3′,5,5′-tetramethylbenzidine, ethylenediaminetetraacetic acid, hexadecyltrimethylammonium bromide and 2,4,6-trinitrobenzene sulfonic acid were purchased from Sigma (St. Louis, MO, USA) and lyophilized. IL-1 and IL-6 were purchased from BD Pharmingen (San Diego, CA, USA). D₂O, Folin Ciocalteau and KBr were purchased from Sigma (St. Louis, MO, USA).

da Nascimento Santos M.S., de Magalhães J.E., Will Castro L.S., de Sousa Pinheiro T., Sabry D.A., Nobre L.T., Lima J.P., Baseia I.G, & Leite E.L. (2014). Effect of Glucans from Caripia montagnei Mushroom on TNBS-Induced Colitis. International Journal of Molecular Sciences, 15(2), 2368-2385.

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Extraction and Characterization of Marine Polysaccharides

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C. antiquata was purchased from ZhanJiang, China. Mammalian heparin (HP), chondroitin sulfate (CS), dermatin sulfate (DS), and plasminogen were obtained from the National Institutes for Food and Drug Control (China). Alkaline protease 2709 (1.2 × 105 U/g), papain (800 U/mg), rabbit plasma, thrombin (300 U/mg), urokinase (50 U/mg), and bovine plasma fibrinogen were obtained from Yuanye Biotechnology Co. Ltd. (Shanghai, China). Activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) kits were purchased from Sun Biotech Co. Ltd. (Shanghai, China). AMBERLITE FPA98 CI macroporous anion exchange resin was purchased from Rohm and Haas (Philadelphia, PA). Standard monosaccharide references (mannose, rhamnose, glucosamine, glucuronic acid, iduronic acid, N-acetylglucosamine, glucose, galactose, arabinose, and fucose) were obtained from Sigma-Aldrich (Darmstadt, Germany). Different molecular weights of heparin standard were purchased from Adhoc International Technology Co., Ltd. (Beijing, China). All other chemicals and solvents used were of analytical grade or HPLC-grade, unless otherwise specified.

Du Z., Jia X., Chen J., Zhou S., Chen J., Liu X., Cao X., Zhong S, & Hong P. (2019). Isolation and Characterization of a Heparin-Like Compound with Potent Anticoagulant and Fibrinolytic Activity from the Clam Coelomactra antiquata. Marine Drugs, 18(1), 6.

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Monosaccharide Analysis of Butyrivibrio Cultures

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The monosaccharides released during Butyrivibrio monoculture and coculture growth were determined using a HPIC method (55 ). Culture samples from the eight time intervals (0, 2, 4, 6, 8, 10, 12 and 16 h) were used for analysis, and the HPIC system used was the DIONEX ICS-5000 system (Thermo Fisher Scientific). Data analysis was performed with Chromeleon software (Thermo Fisher Scientific). Using a two-tailed, unpaired Student's t test, differences between sample means were considered statistically significant if the P value was <0.05. Quantitative analyses were carried out using standard solutions of the monosaccharides arabinose (Sigma-Aldrich), fucose (Sigma-Aldrich), galactose (Sigma-Aldrich), glucose (VWR International Ltd., Poole, UK), rhamnose (VWR), and xylose (VWR).

Palevich N., Kelly W.J., Ganesh S., Rakonjac J, & Attwood G.T. (2019). Butyrivibrio hungatei MB2003 Competes Effectively for Soluble Sugars Released by Butyrivibrio proteoclasticus B316T during Growth on Xylan or Pectin. Applied and Environmental Microbiology, 85(3), e02056-18.

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