Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/13421
Citations
Scopus Web of Science® Altmetric
?
?
Full metadata record
DC FieldValueLanguage
dc.contributor.authorHrmova, M.en
dc.contributor.authorImai, T.en
dc.contributor.authorRutten, S.en
dc.contributor.authorFairweather, J.en
dc.contributor.authorPelosi, L.en
dc.contributor.authorBulone, V.en
dc.contributor.authorDriguez, H.en
dc.contributor.authorFincher, G.en
dc.date.issued2002en
dc.identifier.citationJournal of Biological Chemistry, 2002; 277(33):30102-30111en
dc.identifier.issn0021-9258en
dc.identifier.issn1083-351Xen
dc.identifier.urihttp://hdl.handle.net/2440/13421-
dc.descriptionCopyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.en
dc.description.abstractBarley (1,3)-beta-D-glucan endohydrolases (EC ), inactivated by site-directed mutagenesis of their catalytic nucleophiles, show autocondensation glucosynthetic activity with alpha-laminaribiosyl fluoride and heterocondensation glycosynthetic activity with alpha-laminaribiosyl fluoride and 4'-nitrophenyl beta-D-glucopyranoside. The native enzyme is a retaining endohydrolase of the family 17 group and catalyzes glycosyl transfer reactions at high substrate concentrations. Catalytic efficiencies (k(cat) K(m)(-1)) of mutants E231G, E231S, and E231A as glycosynthases are 28.9, 0.9, and 0.5 x 10(-4) m(-1) s(-1), respectively. Glycosynthase reactions appear to be processive and proceed with pH optima of 6-8 and yields of up to 75%. Insoluble products formed during the glycosynthase reaction appear as lamellar, hexagonal crystals when observed by electron microscopy. Methylation, NMR, and matrix-assisted laser desorption ionization time-of-flight analyses show that the reaction products are linear (1,3)-beta-D-glucans with a degree of polymerization of 30-34, whereas electron and x-ray diffraction patterns indicate that these (1,3)-beta-D-glucan chains adopt a parallel, triple helical conformation. The (1,3)-beta-D-glucan triple helices are orientated perpendicularly to the plane of the lamellar crystals. The barley (1,3)-beta-D-glucan glycosynthases have considerable potential for tailored and high efficiency synthesis of (1,3)-beta-D-linked oligo- and polysaccharides, some of which could have immunomodulating activity, or for the coupling of (1,3)-beta-D-linked glucosyl residues onto other oligosaccharides or glycoproteins.en
dc.description.statementofresponsibilityMaria Hrmova, Tomoya Imai, Simon J. Rutten, Jon K. Fairweather, Ludovic Pelosi, Vincent Bulone, Hugues Driguez, and Geoffrey B. Fincheren
dc.language.isoenen
dc.publisherAmer Soc Biochemistry Molecular Biology Incen
dc.subjectHordeum; Glucans; Glycoside Hydrolases; beta-Glucans; DNA Primers; Microscopy, Electron; Crystallization; Magnetic Resonance Spectroscopy; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Mutagenesis, Site-Directed; Base Sequence; Carbohydrate Sequence; Catalysisen
dc.titleMutated Barley (1,3)-beta-D-Glucan Endohydrolases Synthesize Crystalline (1,3)-beta-D-Glucansen
dc.typeJournal articleen
dc.identifier.rmid0020020632en
dc.identifier.doi10.1074/jbc.M203971200en
dc.identifier.pubid60262-
pubs.library.collectionAgriculture, Food and Wine publicationsen
pubs.verification-statusVerifieden
pubs.publication-statusPublisheden
dc.identifier.orcidHrmova, M. [0000-0002-3545-0605]en
dc.identifier.orcidBulone, V. [0000-0002-9742-4701]en
Appears in Collections:Agriculture, Food and Wine publications

Files in This Item:
There are no files associated with this item.


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.