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• W2007604144 abstract "Carbohydrate-active enzymes have multiple biological roles and industrial applications. Advances in genome and transcriptome sequencing together with associated bioinformatics tools have identified vast numbers of putative carbohydrate-degrading and -modifying enzymes including glycoside hydrolases and lytic polysaccharide monooxygenases. However, there is a paucity of methods for rapidly screening the activities of these enzymes. By combining the multiplexing capacity of carbohydrate microarrays with the specificity of molecular probes, we have developed a sensitive, high throughput, and versatile semiquantitative enzyme screening technique that requires low amounts of enzyme and substrate. The method can be used to assess the activities of single enzymes, enzyme mixtures, and crude culture broths against single substrates, substrate mixtures, and biomass samples. Moreover, we show that the technique can be used to analyze both endo-acting and exo-acting glycoside hydrolases, polysaccharide lyases, carbohydrate esterases, and lytic polysaccharide monooxygenases. We demonstrate the potential of the technique by identifying the substrate specificities of purified uncharacterized enzymes and by screening enzyme activities from fungal culture broths. Carbohydrate-active enzymes have multiple biological roles and industrial applications. Advances in genome and transcriptome sequencing together with associated bioinformatics tools have identified vast numbers of putative carbohydrate-degrading and -modifying enzymes including glycoside hydrolases and lytic polysaccharide monooxygenases. However, there is a paucity of methods for rapidly screening the activities of these enzymes. By combining the multiplexing capacity of carbohydrate microarrays with the specificity of molecular probes, we have developed a sensitive, high throughput, and versatile semiquantitative enzyme screening technique that requires low amounts of enzyme and substrate. The method can be used to assess the activities of single enzymes, enzyme mixtures, and crude culture broths against single substrates, substrate mixtures, and biomass samples. Moreover, we show that the technique can be used to analyze both endo-acting and exo-acting glycoside hydrolases, polysaccharide lyases, carbohydrate esterases, and lytic polysaccharide monooxygenases. We demonstrate the potential of the technique by identifying the substrate specificities of purified uncharacterized enzymes and by screening enzyme activities from fungal culture broths. Almost all plant cells are encased in a cell wall composed primarily of complex polysaccharides (1Albersheim P. Darvill A. Roberts K. Sederoff R. Staehelin A. Plant Cell Walls: from Chemistry to Biology. Garland Science, New York2011: 37-47Google Scholar). Load-bearing components, principally cellulose and hemicelluloses, are embedded in a matrix of pectic polysaccharides, and in mature secondary walls, further reinforcement is provided by the phenolic polymer lignin (2Carpita N.C. Gibeaut D.M. Structural models of primary cell walls in flowering plants: consistency of molecular structure with the physical properties of the walls during growth.Plant J. 1993; 3: 1-30Crossref PubMed Scopus (2869) Google Scholar, 3Cosgrove D.J. Growth of the plant cell wall.Nat. Rev. Mol. Cell Biol. 2005; 6: 850-861Crossref PubMed Scopus (2262) Google Scholar4Vanholme R. Demedts B. Morreel K. Ralph J. Boerjan W. Lignin biosynthesis and structure.Plant Physiol. 2010; 153: 895-905Crossref PubMed Scopus (1621) Google Scholar). Cell walls play crucial roles in plant growth and development. They provide structural support, are physical defensive barriers against biotic and abiotic stress, and are a source of oligosaccharide signaling molecules (2Carpita N.C. Gibeaut D.M. Structural models of primary cell walls in flowering plants: consistency of molecular structure with the physical properties of the walls during growth.Plant J. 1993; 3: 1-30Crossref PubMed Scopus (2869) Google Scholar, 5Malinovsky F.G. Fangel J.U. Willats W.G. The role of the cell wall in plant immunity.Front. Plant Sci. 2014; 5: 178Crossref PubMed Scopus (304) Google Scholar, 6Ridley B.L. Neill M.A. Mohnen D. Pectins: structure, biosynthesis, and oligogalacturonide-related signaling.Phytochemistry. 2001; 57: 929-967Crossref PubMed Scopus (1500) Google Scholar). Collectively, plant cell walls are also the largest source of biomass on earth and have received considerable attention in the context of attempts to produce second generation biofuels and chemicals from lignocellulosic feedstocks as an alternative renewable source to fossil fuels (7Jørgensen H. Kristensen J.B. Felby C. Enzymatic conversion of lignocellulose into fermentable sugars: challenges and opportunities.Biofuels Bioprod. Biorefining. 2007; 1: 119-134Crossref Scopus (872) Google Scholar, 8Pauly M. Keegstra K. Cell-wall carbohydrates and their modification as a resource for biofuels.Plant J. 2008; 54: 559-568Crossref PubMed Scopus (610) Google Scholar9Himmel M.E. Bayer E.A. Lignocellulose conversion to biofuels: current challenges, global perspectives.Curr. Opin. Biotechnol. 2009; 20: 316-317Crossref PubMed Scopus (91) Google Scholar). A vast repertoire of cell wall-degrading enzymes, notably glycoside hydrolases (GHs), 3The abbreviations used are: GHglycoside hydrolaseLPMOlytic polysaccharide monooxygenaseHGhomogalacturonanAZCLazurine cross-linkedCBMcarbohydrate binding moduleCDTAdiaminocyclohexanetetraacetic acidHPAEChigh performance anion-exchange chromatographyDEdegree of methyl esterificationCAZycarbohydrate-active enzymes database. are produced by microbes, and these are vital for strategies aimed at the deconstruction of cell wall-based feedstocks used for biorefining (10Flint H.J. Bayer E.A. Plant cell wall breakdown by anaerobic microorganisms from the mammalian digestive tract.Ann. N.Y. Acad. Sci. 2008; 1125: 280-288Crossref PubMed Scopus (148) Google Scholar, 11Dashtban M. Schraft H. Qin W. Fungal bioconversion of lignocellulosic residues; opportunities & perspectives.Int. J. Biol. Sci. 2009; 5: 578-595Crossref PubMed Scopus (537) Google Scholar12van den Brink J. de Vries R.P. Fungal enzyme sets for plant polysaccharide degradation.Appl. Microbiol. Biotechnol. 2011; 91: 1477-1492Crossref PubMed Scopus (402) Google Scholar). Carbohydrate-degrading and -modifying enzymes are also extensively used in the production of paper, textiles, detergents, feed, and food (13Beg Q.K. Kapoor M. Mahajan L. Hoondal G.S. Microbial xylanases and their industrial applications: a review.Appl. Microbiol. Biotechnol. 2001; 56: 326-338Crossref PubMed Scopus (1081) Google Scholar, 14Kirk O. Borchert T.V. Fuglsang C.C. Industrial enzyme applications.Curr. Opin. Biotechnol. 2002; 13: 345-351Crossref PubMed Scopus (1034) Google Scholar). Endogenous carbohydrate-degrading and -modifying enzymes are central to many plant development processes. For example, GHs are used to cleave polysaccharides during organ abscission, fruit softening, germination, and plant/microbe interactions. Also, plants typically produce methyl- and acetylesterases that are used for fine-tuning the in muro functionalities of cell wall components to suit local requirements (15Wolf S. Mouille G. Pelloux J. Homogalacturonan methyl-esterification and plant development.Mol. Plant. 2009; 2: 851-860Abstract Full Text Full Text PDF PubMed Scopus (307) Google Scholar, 16Franková L. Fry S.C. Biochemistry and physiological roles of enzymes that ’cut and paste’ plant cell-wall polysaccharides.J. Exp. Bot. 2013; 64: 3519-3550Crossref PubMed Scopus (142) Google Scholar). glycoside hydrolase lytic polysaccharide monooxygenase homogalacturonan azurine cross-linked carbohydrate binding module diaminocyclohexanetetraacetic acid high performance anion-exchange chromatography degree of methyl esterification carbohydrate-active enzymes database. Techniques for mining genomes and metagenomes have developed rapidly in recent years, and so have medium and high throughput strategies for cloning and expressing recombinant enzymes. Furthermore, bioinformatics resources and associated depositories, such as the carbohydrate-active enzymes database (CAZy) (17Cantarel B.L. Coutinho P.M. Rancurel C. Bernard T. Lombard V. Henrissat B. The Carbohydrate-Active EnZymes database (CAZy): an expert resource for glycogenomics.Nucleic Acids Res. 2009; 37: D233-D238Crossref PubMed Scopus (4114) Google Scholar) have expanded greatly. However, there are considerable challenges inherent in the exploitation of microbial enzyme diversity for industrial purposes, and the empirical determination of enzyme activities has now become a serious bottleneck. For example, it is estimated that, using current methods, we can safely predict the activities of no more than 4% of the proteins within the CAZy (18Lombard V. Golaconda Ramulu H. Drula E. Coutinho P.M. Henrissat B. The carbohydrate-active enzymes database (CAZy) in 2013.Nucleic Acids Res. 2014; 42: D490-D495Crossref PubMed Scopus (4115) Google Scholar). Although numerous methods are available for monitoring enzyme activities, they generally have some limitations. Well established techniques based on the analysis of the oligomeric fragments produced by enzymatic hydrolysis, such as chromatography combined with mass spectrometry, are available (19Peña M.J. Tuomivaara S.T. Urbanowicz B.R. O’Neill M.A. York W.S. Methods for structural characterization of the products of cellulose- and xyloglucan-hydrolyzing enzymes.Methods Enzymol. 2012; 510: 121-139Crossref PubMed Scopus (35) Google Scholar). These approaches are powerful; however, they are labor-intensive and generally not suitable for high throughput screening. Methods based on the measurement of reducing sugars such as the 3,5-dinitrosalicylic acid (20Miller G.L. Use of dinitrosalicylic acid reagent for determination of reducing sugar.Anal. Chem. 1959; 31: 426-428Crossref Scopus (22220) Google Scholar) and the Nelson-Somogyi (21Somogyi M. Notes on sugar determination.J. Biol. Chem. 1952; 195: 19-23Abstract Full Text PDF PubMed Google Scholar) assays are widely used for rapidly screening GH activities. However, these assays lack polysaccharide specificity because reducing ends from different polysaccharides cannot be distinguished. Chromogenic polysaccharide substrates, such as azurine cross-linked (AZCL) and azo-dyed polymers, are extensively used to screen endo-acting GHs, but the current range of available substrates is limited, and the nature of the chemical modification of these polysaccharides renders them unsuitable substrates for other types of enzymes including esterases and exo-acting GHs. Exo-acting enzymes can be analyzed with para-nitrophenyl substrates, but these compounds can usually only provide information about the degradation of one substrate per reaction and are not effective substrates for all exo-acting enzymes (22Honda Y. Kitaoka M. A family 8 glycoside hydrolase from Bacillus halodurans C-125 (BH2105) is a reducing end xylose-releasing exo-oligoxylanase.J. Biol. Chem. 2004; 279: 55097-55103Abstract Full Text Full Text PDF PubMed Scopus (80) Google Scholar, 23Lagaert S. Van Campenhout S. Pollet A. Bourgois T.M. Delcour J.A. Courtin C.M. Volckaert G. Recombinant expression and characterization of a reducing-end xylose-releasing exo-oligoxylanase from Bifidobacterium adolescentis.Appl. Environ. Microbiol. 2007; 73: 5374-5377Crossref PubMed Scopus (40) Google Scholar24Lagaert S. Pollet A. Delcour J.A. Lavigne R. Courtin C.M. Volckaert G. Characterization of two β-xylosidases from Bifidobacterium adolescentis and their contribution to the hydrolysis of prebiotic xylooligosaccharides.Appl. Microbiol. Biotechnol. 2011; 92: 1179-1185Crossref PubMed Scopus (43) Google Scholar). There is therefore a pressing need for the development of new technology for the rapid screening of GHs and other carbohydrate-active enzymes. We describe the development of a semiquantitative method that combines the multiplexing capacity of robotically produced microarrays with the specificity of monoclonal antibodies (mAbs) and carbohydrate binding modules (CBMs). The method is versatile and can be used to assess enzyme activities against both pure substrates and complex mixtures of cell wall components. In addition to GH activities, the technique can be used for the analysis of other polysaccharide-modifying enzymes including carbohydrate esterases, polysaccharide lyases, and lytic polysaccharide monooxygenases. We demonstrate the applicability of the technique for identifying the activities of novel putative enzymes and for screening crude fungal culture broths. Commercial enzymes from Megazyme (Bray, Ireland) used in this work are depicted in Table 1. The enzyme mixture Cellic Ctec2; the purified enzymes eNZ1 (endo-1,4-β-xylanase), eNZ2 (endo-1,4-β-xylanase), eNZ3 (endo-1,4-β-glucanase), and eNZ4 (endo-1,4-β-mannanase) (for details, see Table 1); the 24 crude culture broths obtained from the fungi Gibberella zeae, Poronia punctata, Fusarium oxysporum, Magnaporthe grisea, Ustilago maydis, and Deconica inquilina grown under four different conditions (samples were grown for 5 days at pH 6.5–9.0 in two different growth media (yeast extract-peptone-glucose and FG-4)) (Fig. 6); and the crude recombinant Aspergillus nidulans endo-1,4-β-mannanase culture broth (Fig. 9B) were provided by Novozymes (Bagsværd, Denmark). NcLPMO9C was supplied by Vincent G. H. Eijsink.TABLE 1Characterized enzymes and enzyme mixture used in this studyCodeEnzyme nameSourceMicroorganismCatalogue no.N/Aα-l-ArabinofuranosidaseMegazymeBifidobacterium sp.E-AFAM2eCELCellulaseMegazymeTrichoderma longibrachiatumE-CELTRN/AFeruloyl esteraseMegazymeRumen microorganismE-FAERUeGLCEndo-1,3-β-glucanaseMegazymeTrichoderma sp.E-LAMSEN/AExo-1,3-β-glucanaseMegazymeN/AN/AN/AEndo-1,3(4)-β-glucanaseMegazymeBacillus subtilisE-LICHNeMANEndo-1,4-β-mannanaseMegazymeCellvibrio japonicusE-BMACJN/APectate lyaseMegazymeAspergillus sp.E-PECLYePOLPolygalacturonase M2MegazymeAspergillus aculeatusE-PGALUSPeXYLEndo-1,4-β-xylanase M4MegazymeAspergillus nigerE-XYAN4N/AExo-1,4-β-xylosidaseMegazymeBacillus pumilusE-BXSEBPeXGXyloglucanase (GH5)MegazymePaenibacillus sp.E-XEGPeNZ1Endo-1,4-β-xylanase (GH10)NovozymesAspergillus aculeatusN/AeNZ2Endo-1,4-β-xylanase (GH11)NovozymesThermomyces lanuginosusN/AeNZ3Endo-1,4-β-glucanase (GH5)NovozymesAspergillus aculeatusN/AeNZ4Endo-1,4-β-mannanase (GH5)NovozymesFungal origin (proprietary)N/AN/ACellic Ctec2NovozymesMultienzyme productN/AN/ANcLPMO9CV. G. H. EijsinkNeurospora crassaN/A Open table in a new tab FIGURE 9Resolving activities against specific epitopes within complex substrate mixtures. A, three enzymes (xyloglucanase at 0. 001 unit/ml, endo-1,3-β-glucanase at 0.01 unit/ml, and polygalacturonase at 0.01 unit/ml) and an enzyme mixture (Cellic Ctec2 used at 1:20 dilution) listed to the right were tested against the two mixtures of substrates shown to the left, and activities were detected with the probes listed at the top of the heat map. The -fold change values in the heat map show that individual activities can be resolved from the substrate mixtures by the specific binding of the probes. Reactions were performed for 1 h at 40 °C. B, similarly, a crude recombinant A. nidulans endo-1,4-β-mannanase culture broth was also tested against a mixture of polysaccharide substrates. Note that side activities progressively diminished with decreasing broth concentration (shown to the right). Reactions were performed for 1 h at 30 °C. A and B, substrate mixtures contained 0.1 mg/ml (per polysaccharide). RGI, rhamnogalacturonan I; AX, arabinoxylan. Details of enzymes and probes used are provided in TABLE 1, TABLE 2, respectively.View Large Image Figure ViewerDownload Hi-res image Download (PPT) Pseudacanthotermes militaris proteins were obtained by the amplification of metagenomic DNA fragments by polymerase chain reaction using Phusion high fidelity DNA polymerase (New England Biolabs). The amplicons were cloned as described (25Vincentelli R. Cimino A. Geerlof A. Kubo A. Satou Y. Cambillau C. High-throughput protein expression screening and purification in Escherichia coli.Methods. 2011; 55: 65-72Crossref PubMed Scopus (75) Google Scholar), and recombinant proteins Pm12, Pm23, and Pm25 were expressed in fusion with a polyhistidine tag, whereas Pm83, Pm84, and Pm85 were produced in fusion with a polyhistidine-thioredoxin tag. Plasmids were used to transform Escherichia coli Rosetta DE3 (Merck Millipore, Germany). For protein expression, the autoinducing medium ZYP-5052 (prepared as described in Ref. 26Studier F.W. Protein production by auto-induction in high-density shaking cultures.Protein Expr. Purif. 2005; 41: 207-234Crossref PubMed Scopus (4109) Google Scholar) supplemented with chloramphenicol (34 μg/ml) was used. Cells were incubated at 37 °C for 5 h and at 17 °C overnight in baffled flasks until an A600 nm of ∼11 was reached. Immobilized metal ion affinity chromatography was performed following the protocol described previously (27Bastien G. Arnal G. Bozonnet S. Laguerre S. Ferreira F. Fauré R. Henrissat B. Lefèvre F. Robe P. Bouchez O. Noirot C. Dumon C. O’Donohue M. Mining for hemicellulases in the fungus-growing termite Pseudacanthotermes militaris using functional metagenomics.Biotechnol. Biofuels. 2013; 6: 78Crossref PubMed Scopus (54) Google Scholar) for purification of the recombinant proteins. The concentration of purified proteins was determined using a NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific) and applying the Beer-Lambert law. The theoretical extinction coefficient and molecular weight values were determined using the ProtParam service. Sequence data can be found in GenBankTM under the following accession numbers: CCO21106.1 (Pm12), CCO21108.1 (Pm23), CCO21110.1 (Pm25), CCO21640.1 (Pm83), CCO21632.1 (Pm84), and CCO21658.1 (Pm85). Fungal broth supernatants (Fig. 13) were prepared as follows. Baffled Erlenmeyer flasks (500 ml) with 200 ml of Czapek Dox-based medium with apple pomace (35.3 mm NaNO3, 5.7 mm K2HPO4, 6.7 mm KCl, 2 mm MgSO4, 1 g of Triton-X (0.1%), 1 ml of trace metal solution (36 mm FeSO4, 11.5 mm ZnSO4, 17.5 mm CuSO4) and 20 g of ground apple pomace in 1000 ml of Milli-Q water) were autoclaved and afterward inoculated with ¼ agar plate of Trametes versicolor (DSM-11269, Deutsche Sammlung von Mikroorganismen und Zellkulturen, Germany), 1 × 104 spores of Colletotrichum acutatum (isolate SA 0-1, proprietary), or 1 × 104 spores of Penicillium expansum (isolate IK 2020, proprietary) or mock-inoculated as a negative control. The flasks were incubated at 25 °C with shaking at 150 rpm for 7 days in darkness. Samples were collected from day 2 to 7 under sterile conditions and centrifuged twice for 20 min at 4000 rpm at 4 °C to separate spores from supernatants. Supernatants were collected, frozen in liquid nitrogen, and stored at −80 °C until use. Commercial enzymes were analyzed within their recommended pH range using the following buffers: 0.1 m sodium acetate buffer for pH values 4.0–6.5 and 0.1 m sodium phosphate buffer for pH values 7.0–8.0 except for pH 2–10 analysis where 0.1 m Britton-Robinson buffer was used (Fig. 7). The rest of the samples were analyzed using the following buffers: the eNZ1–4 in 50 mm sodium acetate at pH 5.5; the 24 crude broths (Fig. 6) and the recombinant A. nidulans endo-1,4-β-mannanase broth (Fig. 9B) in 0.1 m sodium acetate, pH 5.5 and pH 5.0, respectively; NcLPMO9C in 0.1 m sodium phosphate, pH 8.0; P. militaris proteins in 0.1 m sodium acetate, pH 6.5; and fungal broths (Fig. 13) in 0.2 m sodium acetate, pH 6. Enzyme concentrations used are given in the figure legends. The following defined polysaccharides were used in this work: arabinan (sugar beet), arabinoxylan (wheat flour), β-glucan (barley), β-glucan lichenan (icelandic moss), galactomannan (carob), glucomannan (konjac), pachyman (1,3-β-d-glucan), pectic galactan (lupin), and xyloglucan (tamarind) from Megazyme; gum arabic (acacia tree), hydroxyethylcellulose, and xylan (beechwood) from Sigma-Aldrich; pectin with degrees of methyl esterification (DEs) of 11, 16, and 81% (lime) from DuPont Nutrition Biosciences (Brabrand, Denmark); and feruloylated arabinoxylan (wheat flour) from Institut National de la Recherche Agronomique (Nantes, France). Polysaccharide substrates used in Fig. 10 were extracted as described (28Moller I. Sørensen I. Bernal A.J. Blaukopf C. Lee K. Øbro J. Pettolino F. Roberts A. Mikkelsen J.D. Knox J.P. Bacic A. Willats W.G. High-throughput mapping of cell-wall polymers within and between plants using novel microarrays.Plant J. 2007; 50: 1118-1128Crossref PubMed Scopus (256) Google Scholar). Briefly, cell wall polysaccharides were extracted from the alcohol-insoluble residues of Arabidopsis thaliana (rosette), Salix alba (willow with bark), Zea mays (corn stover, whole), Hordeum vulgare (barley, stem, and leaves), and Equisetum arvense (horsetail, stem, and leaves). Sequential extractions with 50 mm CDTA and 4 m sodium hydroxide (NaOH) with 1% (v/v) NaBH4 were performed. Extracts were neutralized to pH 6–7 with glacial acetic acid. Defined polysaccharides were dissolved in deionized water to 4 mg/ml except for pachyman, which was dissolved in 4 m NaOH. Afterward both single polysaccharides and polysaccharide mixtures were prepared by diluting them in printing buffer (55.2% glycerol, 44% water, 0.8% Triton X-100). Substrate concentrations used (generally 0.1 mg/ml) are specified in the figure legends. Substrate mixtures contained each polysaccharide at the same concentration. Additionally, the pH of pachyman solution was neutralized with glacial acetic acid considering final concentration. Regarding the plant extracted polysaccharides (Fig. 10), a 2-fold dilution followed by a 5-fold dilution was performed in printing buffer. Substrate solutions (defined polysaccharides or plant extracts) were added into wells of 384-microwell plates (PP microplate, V-shape, catalogue number 781280, Greiner Bio-One), and afterward the enzymatic samples were added (substrate/sample, 1:1, v/v) to a final volume of 20 μl/well. Controls were prepared for each substrate solution without enzyme, keeping all other conditions identical. Each reaction was performed in triplicate (except plant extracts that had duplicates at two different concentrations). The filled plates were incubated (Ecotron, INFORS HT, Switzerland) at 100 rpm for a specific time and temperature (specified in the figure legends) followed by 10 min at 80 °C. After spinning down the plates for 10 min at 4000 rpm, the plate content was printed at 22 °C and 55% humidity onto nitrocellulose membrane with a pore size of 0.45 μm (Whatman) using a microarray robot (Sprint, Arrayjet, Roslin, UK). The printed arrays were blocked for 1 h in PBS (140 mm NaCl, 2.7 mm KCl, 10 mm Na2HPO4, 1.7 mm KH2PO4, pH 7.5) with 5% (w/v) low fat milk powder (MPBS). Then arrays were incubated for 2 h with probes (for details, see Table 2) that for the different experiments included anti-rat and anti-mouse mAbs and CBMs (PlantProbes, Leeds, UK; Institut National de la Recherche Agronomique, Nantes, France; BioSupplies, Bundoora, Australia; and NZYTech, Lisbon, Portugal). Probes were diluted 1:10, 1:1000, and 10 μg/ml, respectively, in MPBS. Afterward arrays were washed thoroughly in PBS and incubated for 2 h with anti-rat, anti-mouse, or anti-His tag secondary antibodies conjugated to alkaline phosphatase (Sigma) diluted 1:5000 (anti-rat and anti-mouse) or 1:1500 (anti-His tag) in MPBS. Once washed in PBS and deionized water, microarrays were developed in a solution containing 5-bromo-4-chloro-3-indolylphosphate and nitro blue tetrazolium in alkaline phosphatase buffer (100 mm NaCl, 5 mm MgCl2, 100 mm diethanolamine, pH 9.5).TABLE 2Specificities of monoclonal antibodies and CBMs used in this studyProbeRecognized epitopeRef.JIM5Partially methyl-esterified/de-esterified HG44Clausen M.H. Willats W.G. Knox J.P. Synthetic methyl hexagalacturonate hapten inhibitors of anti-homogalacturonan monoclonal antibodies LM7, JIM5 and JIM7.Carbohydr. Res. 2003; 338: 1797-1800Crossref PubMed Scopus (256) Google ScholarJIM7Partially methyl-esterified HG44Clausen M.H. Willats W.G. Knox J.P. Synthetic methyl hexagalacturonate hapten inhibitors of anti-homogalacturonan monoclonal antibodies LM7, JIM5 and JIM7.Carbohydr. Res. 2003; 338: 1797-1800Crossref PubMed Scopus (256) Google ScholarLM18Partially methyl-esterified/de-esterified HG45Verhertbruggen Y. Marcus S.E. Haeger A. Ordaz-Ortiz J.J. Knox J.P. An extended set of monoclonal antibodies to pectic homogalacturonan.Carbohydr. Res. 2009; 344: 1858-1862Crossref PubMed Scopus (307) Google ScholarLM19Partially methyl-esterified/de-esterified HG45Verhertbruggen Y. Marcus S.E. Haeger A. Ordaz-Ortiz J.J. Knox J.P. An extended set of monoclonal antibodies to pectic homogalacturonan.Carbohydr. Res. 2009; 344: 1858-1862Crossref PubMed Scopus (307) Google ScholarLM20Partially methyl-esterified HG45Verhertbruggen Y. Marcus S.E. Haeger A. Ordaz-Ortiz J.J. Knox J.P. An extended set of monoclonal antibodies to pectic homogalacturonan.Carbohydr. Res. 2009; 344: 1858-1862Crossref PubMed Scopus (307) Google ScholarINRA-RU1Rhamnogalacturonan I backbone46Ralet M.C. Tranquet O. Poulain D. Moïse A. Guillon F. Monoclonal antibodies to rhamnogalacturonan I backbone.Planta. 2010; 231: 1373-1383Crossref PubMed Scopus (94) Google ScholarINRA-RU2Rhamnogalacturonan I backbone46Ralet M.C. Tranquet O. Poulain D. Moïse A. Guillon F. Monoclonal antibodies to rhamnogalacturonan I backbone.Planta. 2010; 231: 1373-1383Crossref PubMed Scopus (94) Google ScholarLM5(1→4)-β-d-Galactan47Jones L. Seymour G.B. Knox J.P. Localization of pectic galactan in tomato cell walls using a monoclonal antibody specific to (1→4)-β-D-galactan.Plant Physiol. 1997; 113: 1405-1412Crossref PubMed Scopus (367) Google ScholarLM6(1→5)-α-l-Arabinan48Willats W.G. Marcus S.E. Knox J.P. Generation of monoclonal antibody specific to (1→5)-α-L-arabinan.Carbohydr. Res. 1998; 308: 149-152Crossref PubMed Scopus (317) Google ScholarLM12Feruloylated polymers49Pedersen H.L. Fangel J.U. McCleary B. Ruzanski C. Rydahl M.G. Ralet M.C. Farkas V. von Schantz L. Marcus S.E. Andersen M.C. Field R. Ohlin M. Knox J.P. Clausen M.H. Willats W.G. Versatile high resolution oligosaccharide microarrays for plant glycobiology and cell wall research.J. Biol. Chem. 2012; 287: 39429-39438Abstract Full Text Full Text PDF PubMed Scopus (179) Google ScholarLM21Heteromannan50Marcus S.E. Blake A.W. Benians T.A. Lee K.J. Poyser C. Donaldson L. Leroux O. Rogowski A. Petersen H.L. Boraston A. Gilbert H.J. Willats W.G. Knox J.P. Restricted access of proteins to mannan polysaccharides in intact plant cell walls.Plant J. 2010; 64: 191-203Crossref PubMed Scopus (191) Google ScholarBS-400-4(1→4)-β-d-Mannan51Pettolino F.A. Hoogenraad N.J. Ferguson C. Bacic A. Johnson E. Stone B.A. A (1→4)-β-mannan-specific monoclonal antibody and its use in the immunocytochemical location of galactomannans.Planta. 2001; 214: 235-242Crossref PubMed Scopus (58) Google ScholarBS-400-2(1→3)-β-d-Glucan52Meikle P.J. Bonig I. Hoogenraad N.J. Clarke A.E. Stone B.A. The location of (1→3)-β-glucans in the walls of pollen tubes of Nicotiana alata using a (1→3)-β-glucan-specific monoclonal antibody.Planta. 1991; 185: 1-8Crossref PubMed Scopus (193) Google ScholarBS-400-3(1→3)(1→4)-β-d-Glucan53Meikle P.J. Hoogenraad N.J. Bonig I. Clarke A.E. Stone B.A. A (1→3,1→4)-β-glucan-specific monoclonal antibody and its use in the quantitation and immunocyto-chemical location of (1→3,1→4)-β-glucans.Plant J. 1994; 5: 1-9Crossref PubMed Scopus (150) Google ScholarLM15Xyloglucan (XXXG motif)54Marcus S.E. Verhertbruggen Y. Hervé C. Ordaz-Ortiz J.J. Farkas V. Pedersen H.L. Willats W.G. Knox J.P. Pectic homogalacturonan masks abundant sets of xyloglucan epitopes in plant cell walls.BMC Plant Biol. 2008; 8: 60Crossref PubMed Scopus (321) Google ScholarLM25Xyloglucan (XXXG/galactosylated)49Pedersen H.L. Fangel J.U. McCleary B. Ruzanski C. Rydahl M.G. Ralet M.C. Farkas V. von Schantz L. Marcus S.E. Andersen M.C. Field R. Ohlin M. Knox J.P. Clausen M.H. Willats W.G. Versatile high resolution oligosaccharide microarrays for plant glycobiology and cell wall research.J. Biol. Chem. 2012; 287: 39429-39438Abstract Full Text Full Text PDF PubMed Scopus (179) Google ScholarLM10(1→4)-β-d-Xylan55McCartney L. Marcus S.E. Knox J.P. Monoclonal antibodies to plant cell wall xylans and arabinoxylans.J. Histochem. Cytochem. 2005; 53: 543-546Crossref PubMed Scopus (379) Google ScholarLM11(1→4)-β-d-Xylan/arabinoxylan55McCartney L. Marcus S.E. Knox J.P. Monoclonal antibodies to plant cell wall xylans and arabinoxylans.J. Histochem. Cytochem. 2005; 53: 543-546Crossref PubMed Scopus (379) Google ScholarJIM8Arabinogalactan protein56Pennell R.I. Janniche L. Kjellbom P. Scofield G.N. Peart J.M. Roberts K. Developmental regulation of a plasma membrane arabinogalactan protein epitope in oilseed rape flowers.Plant Cell. 1991; 3: 1317-1326Crossref PubMed Scopus (240) Google ScholarJIM13Arabinogalactan protein57Knox J.P. Linstead P.J. Peart J. Cooper C. Roberts K. Developmentally regulated epitopes of cell surface arabinogalactan proteins and their relation to root tissue pattern formation.Plant J. 1991; 1: 317-326Crossref PubMed Scopus (327) Google ScholarCBM3aCellulose (crystalline)58Blake A.W. McCartney L. Flint J.E. Bolam D.N. Boraston A.B. Gilbert H.J. Knox J.P. Understanding the biological rationale for the diversity of cellulose-directed carbohydrate-binding modules in prokaryotic enzymes.J. Biol. Chem. 2006; 281:" @default.
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• W2007604144 cites W102745963 @default.
• W2007604144 cites W1877015315 @default.
• W2007604144 cites W1963655252 @default.
• W2007604144 cites W1971651699 @default.
• W2007604144 cites W1980998830 @default.
• W2007604144 cites W1981273978 @default.
• W2007604144 cites W1982737710 @default.
• W2007604144 cites W1991602629 @default.
• W2007604144 cites W1992221689 @default.
• W2007604144 cites W1993071481 @default.
• W2007604144 cites W2001396309 @default.
• W2007604144 cites W2003604861 @default.
• W2007604144 cites W2006594369 @default.
• W2007604144 cites W2007945994 @default.
• W2007604144 cites W2009290346 @default.
• W2007604144 cites W2009555092 @default.
• W2007604144 cites W2009674410 @default.
• W2007604144 cites W2018108021 @default.
• W2007604144 cites W2019847443 @default.
• W2007604144 cites W2025968524 @default.
• W2007604144 cites W2027426885 @default.
• W2007604144 cites W2053789753 @default.
• W2007604144 cites W2054172648 @default.
• W2007604144 cites W2054796509 @default.
• W2007604144 cites W2055328114 @default.
• W2007604144 cites W2057705279 @default.
• W2007604144 cites W2057892597 @default.
• W2007604144 cites W2060579262 @default.
• W2007604144 cites W2071042312 @default.
• W2007604144 cites W2071618396 @default.
• W2007604144 cites W2082533156 @default.
• W2007604144 cites W2087070794 @default.
• W2007604144 cites W2090600260 @default.
• W2007604144 cites W2095239496 @default.
• W2007604144 cites W2095660130 @default.
• W2007604144 cites W2098695754 @default.
• W2007604144 cites W2102309264 @default.
• W2007604144 cites W2104046482 @default.
• W2007604144 cites W2106246537 @default.
• W2007604144 cites W2106420560 @default.
• W2007604144 cites W2106706785 @default.
• W2007604144 cites W2108929776 @default.
• W2007604144 cites W2121368083 @default.
• W2007604144 cites W2122559203 @default.
• W2007604144 cites W2123788376 @default.
• W2007604144 cites W2134465386 @default.
• W2007604144 cites W2135693122 @default.
• W2007604144 cites W2137645073 @default.
• W2007604144 cites W2139114583 @default.
• W2007604144 cites W2149493030 @default.
• W2007604144 cites W2149864075 @default.
• W2007604144 cites W2154506758 @default.
• W2007604144 cites W2156803450 @default.
• W2007604144 cites W2159080918 @default.
• W2007604144 cites W2162211843 @default.
• W2007604144 cites W2164793821 @default.
• W2007604144 cites W2165194260 @default.
• W2007604144 cites W4231121696 @default.
• W2007604144 cites W4232968793 @default.
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