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Characterization of dextransucrases fromLeuconostoc mesenteroides NRRL B-1299

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Abstract

Leuconostoc mesenteroides NRRL B-1299 dextransucrase was fractionated into soluble (SGT) and insoluble (IGT) enzyme preparations differing by their dextran content. In spite of this, they displayed the same Km for sucrose (10 g/L) and the same activation energy (35 kJ/mol). But the presence of cells and insoluble dextran led to the IGT behaving like an immobilized enzyme: stabilization against thermal denaturation and diffusional limitations at low substrate concentrations were observed. On the other hand, the behavior of SGT was influenced by the presence, in the preparation, of soluble dextran that reduced enzyme inhibition by excess substrate. SGT and IGT present very different pH profiles. In the presence of 4 g/L of soluble dextran, IGT was activated and displayed the same susceptibility to pH as SGT. The activation of IGT was highly dependent on the nature of the acceptor added but also on the pH of the reaction medium. IGT and SGT synthesize both soluble and insoluble polymer containing α(l → 2), α(l → 3), and α(l → 6) linkages. A larger amount of insoluble dextran is elaborated by SGT. The polymer structures, examined by13C NMR spectrometry, revealed that they differ mainly by their α(l → 3) linkage content (from 0 to 11%). This linkage seems to be partly responsible for the dextran insolubility and can be completely eliminated by carrying out the synthesis of soluble polymer at pH 7.4 with SGT.

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Authors and Affiliations

  1. Centre de Bioingénierie Gilbert Durand, UMR CNRS 5504 & Lab. Ass. INRA, I.N.S.A., Complexe Scientifique de Rangueil, 31 077, Toulouse cedex, France

    Marguerite Dols, Magali Remaud-Simeon, René-Marc Willemot & Pierre F. Monsan

  2. CERMAV CNRS, BP 53X, Grenoble, France

    Michel Vignon

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  1. Marguerite Dols
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  2. Magali Remaud-Simeon
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  3. René-Marc Willemot
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  4. Michel Vignon
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  5. Pierre F. Monsan
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Correspondence to Pierre F. Monsan.

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Dols, M., Remaud-Simeon, M., Willemot, RM. et al. Characterization of dextransucrases fromLeuconostoc mesenteroides NRRL B-1299. Appl Biochem Biotechnol 62, 47–59 (1997). https://doi.org/10.1007/BF02787983

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  • DOI: https://doi.org/10.1007/BF02787983

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