Structural analysis of bioengineering α-Δ-Glucan produced by a triple mutant of the glucansucrase GTF180 enzyme from Lactobacillus reuteri strain 180: Generation of (α1→4) linkages in a native (1→3)(1→6)-α-Δ-Glucan
article
Site-directed mutagenesis of the glucansucrase gtf180 gene from Lactobacillus reuteri strain 180 was used to transform the active site region. The β-D-glucan (mEPS-PNNS) produced by the triple mutant V1027P:S1137N: A1139S differed in structure from that of the wild-type β-D-glucan (EPS180). Besides (→-3) and (→6) linkages, as present in EPS180, mEPS-PNNS also contained (→1-4) linkages. Linkage analysis, periodate oxidation, and 1D/2D <sup>1</sup>H NMR spectroscopy of the intact mEPS-PNNS, as well as MS and NMR analysis of oligosaccharides obtained by partial acid hydrolysis of mEPS-PNNS afforded a composite model, which includes all identified structural features. © 2008 American Chemical Society.
Chemicals/CAS: periodate, 15056-35-6; alternansucrase, 100630-46-4; glucan, 9012-72-0, 9037-91-6; glycosyltransferase, 9033-07-2; lactic acid, 113-21-3, 50-21-5; oxygen, 7782-44-7; periodic acid, 10450-60-9, 13444-71-8; alternansucrase, EC 2.4.1.140; Anions; Glucans; Glycosyltransferases, EC 2.4.-; Lactic Acid, 50-21-5; metaperiodate, 7790-28-5; Oxygen, 7782-44-7; Periodic Acid, 10450-60-9
Chemicals/CAS: periodate, 15056-35-6; alternansucrase, 100630-46-4; glucan, 9012-72-0, 9037-91-6; glycosyltransferase, 9033-07-2; lactic acid, 113-21-3, 50-21-5; oxygen, 7782-44-7; periodic acid, 10450-60-9, 13444-71-8; alternansucrase, EC 2.4.1.140; Anions; Glucans; Glycosyltransferases, EC 2.4.-; Lactic Acid, 50-21-5; metaperiodate, 7790-28-5; Oxygen, 7782-44-7; Periodic Acid, 10450-60-9
Topics
Food additivesStructural analysisActive sitesGlucansucraseLinkage analysisPeriodate oxidationSite-directed mutagenesisTriple mutantWild-typeChainsLactobacillus reuterilinkage analysisLactobacillus reuteriAnionsChromatography, Ion ExchangeGlucansGlycosyltransferasesHydrogen-Ion ConcentrationHydrolysisLactic AcidLactobacillus reuteriMagnetic Resonance SpectroscopyMass SpectrometryMutagenesis, Site-DirectedMutationOxygenPeriodic AcidProtein Engineering
TNO Identifier
280038
ISSN
15257797
Source
Biomacromolecules, 9(8), pp. 2251-2258.
Pages
2251-2258
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