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Tamarind xyloglucan

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Tamarind xyloglucan is a polysaccharide derived from the seeds of the tamarind tree. It is a complex carbohydrate composed of a backbone of glucose units with xylose side chains. Tamarind xyloglucan is commonly used in various laboratory applications due to its unique physical and chemical properties.

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Lab products found in correlation

Konjac glucomannan, by Megazyme (3 mentions) Carob galactomannan, by Megazyme (3 mentions) Avicel ph 101, by Merck Group (3 mentions) Barley β glucan, by Megazyme (3 mentions) Wheat arabinoxylan, by Megazyme (2 mentions) Potato galactan, by Megazyme (2 mentions) Guar galactomannan, by Megazyme (2 mentions) Sugar beet arabinan, by Megazyme (2 mentions) Lichenan, by Megazyme (2 mentions) Mixed linkage 1 3 1 4 β glucan mlg from oat, by Megazyme (2 mentions) Starch, by Merck Group (1 mentions) Epoch 2 plate reader, by Agilent Technologies (1 mentions) M1778, by Merck Group (1 mentions) Cellometer cell counter, by Revvity (1 mentions) Microtitre plates, by Thermo Fisher Scientific (1 mentions) A9037, by Merck Group (1 mentions) P xygln, by Megazyme (1 mentions) A7058, by Merck Group (1 mentions) Curdlan, by Megazyme (1 mentions) Carboxymethyl cellulose, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Xyloglucan (21 citations) Tamarind seed xyloglucan (6 citations)

22 protocols using tamarind xyloglucan

1

Raman Spectroscopy of Plant Polysaccharides

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Additionally, to obtain exemplary Raman spectra of individual polysaccharides, commercially available pectins, hemicelluloses and cellulose were studied. High methylated (degree of methylation 80%) pectins were purchased from Herbstreit and Fox (Neuenbürg, Germany), microcrystalline cellulose powder (ca. ~ 20 μm) from Sigma Aldrich and xyloglucan (tamarind, purity >95%) from Megazyme (Bray, Ireland). All the chemicals were used without further purification. Also, the Raman spectrum from the tomato cell wall was collected for comparison. Characteristic bands of the individual polysaccharides from the reference materials were used for the calibration of the Raman imaging method.
Chylińska M., Szymańska-Chargot M, & Zdunek A. (2014). Imaging of polysaccharides in the tomato cell wall with Raman microspectroscopy. Plant Methods, 10, 14.
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2

Culturing Human and Mouse Bacteroides

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Bacteroides ovatus (MDA-HVS BO001) was isolated and cultured from healthy volunteer’s stool samples in a Whitley anaerobic chamber (10% H2, 5% CO2 and 85% N2). Human-derived B. ovatus (ATCC 8483) and human-derived B. theta (ATCC 29148) were purchased from American Type Culture Collection (ATCC). Mouse-derived BT (MDA-JAX BT001) was previously isolated15 (link). Bacterial number was quantified using a Nexcelom Cellometer cell counter with SYTO BC dye and propidium iodide. Bacterial growth experiments were performed in a liquid media, BYEM10, composed of a hybrid of BHI and M10 supplemented with yeast extract as previously described15 (link),45 (link). Bacteria were cultured up to 24 or 48 hours at a starting concentration of 1 × 106 bacteria/ml in BYEM10 broth (pH 7.2) with or without 5 mg/ml of porcine gastric mucin (M1778, Sigma-Aldrich), wheat arabionoxylan (wheat flour; low viscosity; Megazyme), xylan (Beechwood; Megazyme), xyloglucan (Tamarind; Megazyme), or starch (wheat; Sigma-Aldrich). Optical densities (OD600 nm) of bacterial cultures were measured with a BioTek Epoch 2 plate reader.
Hayase E., Hayase T., Mukherjee A., Stinson S.C., Jamal M.A., Ortega M.R., Sanchez C.A., Ahmed S.S., Karmouch J.L., Chang C.C., Flores I.I., McDaniel L.K., Brown A.N., El-Himri R.K., Chapa V.A., Tan L., Tran B.Q., Pham D., Halsey T.M., Jin Y., Tsai W.B., Prasad R., Glover I.K., Ajami N.J., Wargo J.A., Shelburne S., Okhuysen P.C., Liu C., Fowler S.W., Conner M.E., Peterson C.B., Rondon G., Molldrem J.J., Champlin R.E., Shpall E.J., Lorenzi P.L., Mehta R.S., Martens E.C., Alousi A.M, & Jenq R.R. (2023). Bacteroides ovatus alleviates dysbiotic microbiota-induced intestinal graft-versus-host disease. Research Square.
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3

ELISA-based Xyloglucan Quantification

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Initial screening of plant hydroponates and growth media involved enzyme‐linked immunosorbent assays (ELISAs) of materials coated directly onto microtitre ELISA plates (Supporting Information Tables S1, S2). For subsequent ELISA quantification of xyloglucan in hydroponates, shaking culture plant media, solid growth media water extracts and soil alkali extracts, solubilized materials were titrated fivefold onto microtitre plates and incubated in a high salt buffer to ensure efficient microtitre plate well coating overnight before processing with rat monoclonal antibody LM25 to detect xyloglucan (Pedersen et al., 2012). Sample titrations were extended as appropriate to ensure absorbance readings in the range 0–1.0 OD (optical density) which was used to generate xyloglucan equivalents using tamarind xyloglucan (Megazyme International, Bray, Ireland) as a standard. Nitrocellulose prints of solid growth media surfaces were developed with LM25 at 10‐fold dilution followed by anti‐rat immunoglobulin G horseradish peroxidase as previously described (Willats et al., 1998). In situ immunofluorescence analysis of M. polymorpha gemmae was performed with gemmae in agar plugs incubated in volumes of antibody solutions using standard indirect labelling methods (Jackson et al., 2012).
Galloway A.F., Pedersen M.J., Merry B., Marcus S.E., Blacker J., Benning L.G., Field K.J, & Knox J.P. (2017). Xyloglucan is released by plants and promotes soil particle aggregation. The New Phytologist, 217(3), 1128-1136.
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4

Growth Curve Analysis of Xanthomonas citri

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For growth curve analysis, X. citri strains was cultured in LBON medium (1% m/v bacto peptone and 0.5% m/v yeast extract) containing 100 µg mL−1 ampicillin at 30 °C and 200 rpm until mid-exponential phase. Then, the harvested cells were washed once and transferred to the modified minimal medium XVM252 (link) (XVM2m, without sucrose and fructose, containing different sugar sources at a final concentration of 5 mg mL−1), for an initial OD600 nm = 0.01. Growth was monitored for 30 h, at 30 °C, in a SpectraMax M3 Multi-Mode Microplate Reader (Molecular Devices). Four biological replicates were used for each condition. XyGOs used in Xanthomonas growth assays were prepared by incubating 5 mg mL−1 tamarind xyloglucan (Megazyme) with XacXeg74 (4 µg mL−1) at 30 °C for 14 h. The reaction was stopped by heating at 80 °C for 15 min.
Vieira P.S., Bonfim I.M., Araujo E.A., Melo R.R., Lima A.R., Fessel M.R., Paixão D.A., Persinoti G.F., Rocco S.A., Lima T.B., Pirolla R.A., Morais M.A., Correa J.B., Zanphorlin L.M., Diogo J.A., Lima E.A., Grandis A., Buckeridge M.S., Gozzo F.C., Benedetti C.E., Polikarpov I., Giuseppe P.O, & Murakami M.T. (2021). Xyloglucan processing machinery in Xanthomonas pathogens and its role in the transcriptional activation of virulence factors. Nature Communications, 12, 4049.
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5

In vitro Analysis of CBM and Antibody Binding

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In vitro analysis of CBM and antibody binding was carried out using ELISAs as described (12 (link),17 (link)). For antibody/CBM capture ELISAs, 10-fold serial dilutions of 100 μg/ml tamarind xyloglucan (Megazyme) in PBS were coated (100 μl) on to microtitre plates (Nunc, Denmark). LM15 was used at 1:20 dilution of hybridoma cell culture supernatant in MP/PBS. CtCBM3a was used at 10 μg/ml in MP/PBS. Anti-rat-IgG-peroxidase (A9037 Sigma-Aldrich) and anti-his-peroxidase (A7058 Sigma-Aldrich) were used as secondary antibodies for LM15 and CtCBM3a respectively. For competitive-inhibition assessment of recognition of polysaccharides/oligosaccharides microtitre plates were coated with 0.5 μg/ml xyloglucan to act as immobilised antigen. After blocking with MP/PBS and washing, six ten-fold serial dilutions from 50 μg/ml of xyloglucan from tamarind seeds (P-XYGLN Megazyme), xyloglucan heptasaccharide (O-X3G4 Megazyme), guar galactomannan medium viscosity (P-GGMMV Megazyme) and cellohexaose (O-CHE Megazyme) haptens were prepared and 50 μl added to microtritre plate wells. Immediately after, 50 μl of 1:50 dilution of LM15 antibody or CBMs at 20 μg/ml were added to equivalent sets of microtitre plate wells containing the soluble xyloglucan or oligosaccharide haptens and incubated for 1.5 h. Probe binding was determined as described above.
Hernandez-Gomez M.C., Rydahl M.G., Rogowski A., Morland C., Cartmell A., Crouch L., Labourel A., Fontes C.M., Willats W.G., Gilbert H.J, & Knox J.P. (2015). Recognition of xyloglucan by the crystalline cellulose-binding site of a family 3a carbohydrate-binding module. FEBS letters, 589(18), 2297-2303.
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6

Characterization of Diverse Polysaccharides

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Barley beta-glucan (high viscosity), yeast beta-glucan, curdlan, tamarind xyloglucan, konjac glucomannan, carob galactomannan, wheat arabinoxylan, beechwood xylan were purchased from Megazyme International (Bray, Ireland). Laminarin (from Laminaria digitata) was purchased from Sigma Aldrich (St. Louis, MO, USA). Carboxymethyl cellulose was purchased from Acros Organics (Morris Plains, NJ, USA). Hydroxyethyl cellulose was purchased from Amresco (Solon, OH, USA). Xanthan gum was purchased from Spectrum (New Brunswick, NJ, USA). Ulvan (from Ulva sp.) was purchased from Elicityl (Crolles, France).
Tamura K., Foley M.H., Gardill B.R., Dejean G., Schnizlein M., Bahr C.M., Creagh A.L., van Petegem F., Koropatkin N.M, & Brumer H. (2019). Surface glycan binding proteins are essential for cereal beta-glucan utilization by the human gut symbiont Bacteroides ovatus. Cellular and molecular life sciences : CMLS, 76(21), 4319-4340.
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7

Polysaccharide Characterization Protocol

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Fucoidan (F5631), alginate (A7003), laminaran (L9634), carrageenan (C1013), sulfated Dextran (D6001), chondroitin (C4384), oat spelt xylan (X0627), gum Arabic (G9752) and citrus pectin (P9135) were obtained from Sigma-Aldrich. Tamarind xyloglucan, potato galactan, guar galactomannan, sugar beet arabinan and citrus polygalacturonan were obtained from Megazyme International (Bray, Ireland).
Torode T.A., Marcus S.E., Jam M., Tonon T., Blackburn R.S., Hervé C, & Knox J.P. (2015). Monoclonal Antibodies Directed to Fucoidan Preparations from Brown Algae. PLoS ONE, 10(2), e0118366.
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8

Quantification of Xyloglucan Hydrolysis

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To determine any catalytic effect due to truncation of the full-length enzyme [51] (link), the specific activities of BoGH5A and BoGH5A (cat) endo-xyloglucanases were quantified using a bicinchoninic acid (BCA) reducing sugar assay [67] . Each enzyme was used at 0.5 µg/mL. The reactions were conducted in quadruplet in a final volume of 100 µl in 50 mM sodium phosphate buffer (pH 6.0) at 21 ºC for 10 min with 1 mg/mL tamarind xyloglucan (Megazyme, Lot#100403). Reactions were terminated with the addition of an equal volume of BCA working reagent. Color was developed at 80 ºC for 20 min, and the absorbance was read at 563 nm. A series of glucose concentrations (5 to 100 µM) was included in the assay to quantify the amount of reducing ends released. Blank reactions containing only substrate in assay buffer were used to measure the background absorbance generated by reducing ends present on the undigested tamarind xyloglucan. The enzyme concentration used in this assay was not high enough to contribute significantly to the absorbance at 563 nm.
Rodd A.M., Mawhinney W.M, & Brumer H. (2024). A scalable, chromatography-free, biocatalytic method to produce the xyloglucan heptasaccharide XXXG. Biotechnology for biofuels and bioproducts, 17(1).
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9

Enzymatic Activity on Polysaccharides

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The following substrates were employed for exploring the activity of AtAA9A-N and AtAA9B-N: phosphoric acid swollen cellulose (PASC) prepared from Avicel PH-101 (Sigma Aldrich, St. Louis, MO, USA), as described by Wood [48 (link)]; tamarind xyloglucan, cellohexaose (Glc6), cellopentaose (Glc5), Icelandic moss lichenan, and ivory nut mannan, all obtained from Megazyme International Ireland (Wicklow, Ireland); birchwood xylan from Sigma-Aldrich.
Monclaro A.V., Petrović D.M., Alves G.S., Costa M.M., Midorikawa G.E., Miller R.N., Filho E.X., Eijsink V.G, & Várnai A. (2020). Characterization of two family AA9 LPMOs from Aspergillus tamarii with distinct activities on xyloglucan reveals structural differences linked to cleavage specificity. PLoS ONE, 15(7), e0235642.
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10

Xyloglucan Oligosaccharide Labeling Protocol

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XET test papers were prepared as described by Fry (1997) . A piece of Whatman No. 1 filter paper was passed over the surface of 1% (w/v) Tamarind xyloglucan (Megazyme, Ireland) dissolved in 0.5% (w/v) chlorobutanol and left to dry. The dry xyloglucan-coated paper was dipped in 5 µM sulphorhodamine-labelled xyloglucan oligosaccharides (XyGO-SR) dissolved in 75% (v/v) acetone and dried before use. XyGO-SR were prepared by conjugation of xyloglucan oligosaccharides with sulphorhodamine carried out as described by Kosik and Farkas (2008) (link). Xyloglucan-coated papers without XyGO-SR were used as control papers. Tissue prints were made by placing hand-cut 0.5–1.0mm thick pieces of Cuscuta cross-sections on XET test papers or control papers soaked in 50mM Na-acetate, 300mM NaCl (pH 5.5) followed by incubation between two sheets of acetate for 1h. The background on printed papers was removed by washing in water:ethanol:formic acid (1:1:1) for 2h with gentle agitation followed by rinsing in distilled water. Fluorescence micrographs were taken of dried de-stained papers using a SteREO Lumar V12 equipped as described above.
Olsen S., Striberny B., Hollmann J., Schwacke R., Popper Z, & Krause K. (2015). Getting ready for host invasion: elevated expression and action of xyloglucan endotransglucosylases/hydrolases in developing haustoria of the holoparasitic angiosperm Cuscuta. Journal of Experimental Botany, 67(3), 695-708.
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