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  • Title: Structural characterization of bioengineered alpha-D-glucans produced by mutant glucansucrase GTF180 enzymes of Lactobacillus reuteri strain 180.
    Author: van Leeuwen SS, Kralj S, Eeuwema W, Gerwig GJ, Dijkhuizen L, Kamerling JP.
    Journal: Biomacromolecules; 2009 Mar 09; 10(3):580-8. PubMed ID: 19186951.
    Abstract:
    Mutagenesis of specific amino acid residues of the glucansucrase (GTF180) enzyme from Lactobacillus reuteri strain 180 yielded 12 mutant enzymes that produced modified exopolysaccharides (mEPSs) from sucrose. Ethanol-precipitated and purified mEPSs were subjected to linkage analysis, Smith degradation analysis, and 1D/2D (1)H NMR spectroscopy. Comparison of the results with structural data of the previously described wild type EPS180 and triple mutant mEPS-PNNS revealed a broad variation of structural elements between mEPS molecules. The amount of (alpha1-->3) linkages varied from 14-43%, the amount of (alpha1-->4) linkages (not present in the wild type) from 0-12%, and the amount of (alpha1-->6) linkages from 51-86%. The average molecular weight (M(w)) ranged from 9.4 to 32.3 MDa and the degree of branching varied from 8-20%. Using a previously established (1)H NMR structural-reporter-group concept, composite models, that include all identified structural features, were formulated for all mEPS molecules. Variations in the mEPS structures strongly affected the physical properties of the mEPSs.
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