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De-novo synthesis of fructans from sucrose in vitro by a combination of two purified enzymes (sucrose: sucrose 1-fructosyl transferase and fructan: fructan 1-fructosyl transferase) from chicory roots (Cichorium intybus L.)

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Abstract

Although fructans occur widely in several plant families and they have been a subject of investigation for decennia, the mechanism of their biosynthesis is not completely elucidated. We succeeded in purifying a fructan: fructan 1-fructosyl transferase (1-FFT; EC 2.4.1.100) from chicory roots (Cichorium intybus L. var. foliosum cv. Flash). In combination with the purified chicory root sucrose: sucrose 1-fructosyl transferase (1-SST; EC 2.4.1.99), this enzyme synthesized a range of naturally occurring chicory fructans (inulins) from sucrose as the sole substrate. Starting from physiologically relevant sucrose concentrations, inulins up to a degree of polymerization (DP) of about 20 were synthesized in vitro after 96 h at 0°C. Neither 1-SST, nor 1-FFT alone could mediate the observed fructan synthesis. Fructan synthesis in vitro was compared starting from 50, 100 and 200 mM sucrose, respectively. The initiation of (DP > 3)-fructan synthesis was found to be correlated with a certain ratio of 1 kestose to sucrose. The data presented now provide strong evidence to validate the 1-SST/1-FFT model for in-vivo fructan synthesis, at least in the Asteraceae.

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Abbreviations

DP:

degree of polymerization

1-FFT:

fructan: fructan 1-fructosyl transferase

1-SST:

sucrose: sucrose 1-fructosyl transferase

References

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254

    Article  CAS  PubMed  Google Scholar 

  • Cairns AJ (1993) Evidence for the de novo synthesis of fructan by enzymes from higher plants: a reappraisal of the 1-SST/1-FFT model. New Phytol 123: 15–24

    CAS  Google Scholar 

  • Cairns AJ, Ashton JE (1991) The interpretation of in vitro measurements of fructosyl transferase activity: an analysis of patterns of fructosyl transfer by fungal invertase. New Phytol 118: 23–34

    Google Scholar 

  • Cairns AJ, Ashton JE (1993) Species-dependent patterns of fructan synthesis by enzymes from excised leaves of oat, wheat, barley and timothy. New Phytol 124: 381–388

    Google Scholar 

  • Cairns AJ, Ashton JE (1994) Fructan biosynthesis in excised leaves of Lolium temulentum L. VI. Optimization and stability of enzymatic fructan synthesis. New Phytol 126: 3–10

    Google Scholar 

  • Chatterton NJ, Harrison PA, Thornley WR, Bennett JH (1993) Separation and quantification of fructan (inulin) oligomers by anion exchange chromatography. In: Fuchs A (ed) Inulin and inulin-containing crops. Elsevier, Amsterdam, pp 93–99

    Google Scholar 

  • Chen JQ, Black CC (1992) Biochemical and immunological properties of alkaline invertase isolated from sprouting soybean hypocotyls. Arch Biochem Biophys 2951: 61–69

    Google Scholar 

  • Darwen CWE, John P (1989) Localisation of the enzymes of fructan metabolism in vacuoles isolated by a mechanical method from tubers of Jerusalem artichoke (Helianthus tuberosus L.). Plant Physiol 89: 658–663

    Google Scholar 

  • Edelman J, Jefford TG (1968) The mechanism of fructan metabolism in higher plants as exemplified in Helianthus tuberosus. New Phytol 67: 517–531

    Google Scholar 

  • Fuchs A (1991) Current and potential food and non-food applications of fructans. Biochem Soc Trans 19: 555–560

    Google Scholar 

  • Hendry G (1993) Evolutionary origins and natural functions of fructans. A climatological, biogeographic and mechanistic appraisal. New Phytol 123: 3–14

    Google Scholar 

  • Hidaka H, Hirayama M, Yamada K (1991) Fructooligosaccharides. Enzymatic preparation and biofunctions. J Carbohydr Chem 10: 509–522

    Google Scholar 

  • Meier H, Reid JSG (1982) Reserve polysaccharides other than starch in higher plants. In: Loewus FA, Tanner W (eds) Encyclopedia of plant physiology (NS), Plant carbohydrates. Springer-Verlag, Berlin, pp 418–471

    Google Scholar 

  • Penson SP, Cairns AJ (1994) Fructan biosynthesis in excised leaves of wheat (Triticum aestivum L.): a comparison of de novo synthesis in vivo and in vitro. New Phytol 128: 395–402

    Google Scholar 

  • Shiomi N, Onodera S, Chatterton NJ, Harrison PA (1991) Separation of fructo-oligosaccharide isomers by anion-exchange chromatography. Agric Biol Chem 55: 1427–1428

    Google Scholar 

  • Van den Ende W, Van Laere A (1993) Purification and properties of an invertase with sucrose: sucrose fructosyltransferase (1-SST) activity from the roots of Cichorium intybus L. New Phytol 123: 31–37

    Google Scholar 

  • Van den Ende W, Van Laere A (1995) Purification and properties of a neutral invertase from the roots of Cichorium intybus. Physiol Plant 93: 241–248

    Google Scholar 

  • Van den Ende W, Van Wonterghem D, Verhaert P, Dewil E, Van Laere A (1996a) Purification and characterization of fructan: fructan fructosyl transferase from chicory roots (Cichorium intybus L.). Planta, in press

  • Van den Ende W, Van Wonterghem D, Verhaert P, Dewil E, De Loof A, Van Laere A (1996b) Purification and characterization of 1-SST, the key enzyme initiating fructan biosynthesis in young chicory roots (Cichorium intybus L.). Physiologia Plantarum, in press

  • Wiemken A, Frehner M, Keller F, Wagner W (1986) Fructan metabolism, enzymology and compartmentation. Curr Top Plant Biochem Physiol 5: 17–37

    Google Scholar 

  • Wiemken A, Sprenger N, Boller T (1995) Fructans — an extension of sucrose by sucrose. In: Pontis HG, Salerno GL, Echeverria EJ (eds) Current topics in plant physiology, vol 14: Sucrose metabolism, biochemistry, physiology and molecular biology. ASPP, Rockville, pp 179–189

    Google Scholar 

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The authors thank E. Nackaerts for valuable technical assistance. W. Van den Ende is grateful to the National Fund for Scientific Research (NFSR Belgium) for giving a grant for research assistants.

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Van den Ende, W., Van Laere, A. De-novo synthesis of fructans from sucrose in vitro by a combination of two purified enzymes (sucrose: sucrose 1-fructosyl transferase and fructan: fructan 1-fructosyl transferase) from chicory roots (Cichorium intybus L.). Planta 200, 335–342 (1996). https://doi.org/10.1007/BF00200301

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