Summary
A new strain, Claviceps purpurea 88-EP-47, with high invertase activity was selected. Free and Calginate immobilized cultures of this strain were used for fructosylation of ergot alkaloids. By bioconversion from their aglycones, elymoclavine-O-β-d-fructofuranosyl(2→1)-O-β-d-fructofuranoside, and elymoclavine-O-β-d-fructoside, the respective fructosides of chanoclavine, lysergol and dihydro-lysergol monofructosides were obtained. These substances are formed by β-d-fructofuranosidase present in Claviceps cells. The bioconversion activity of the enzyme system (fructose transfer) is strongly dependent on pH, substrate (sucrose) concentration and the developmental profile of invertase activity. The pH optimum for elymoclavine fructosylation is 6.5, for chanoclavine 5.7, and the optimal sucrose concentration is 75 g/l. Fifteen-day-old production cultures had the best glycosylation activities. Fructosylation of alkaloids can be stimulated in production cultures of C. purpurea or C. fusiformis forming elymoclavine or chanoclavine by a pH shift to 6.5 at the end of the production phase. Glycosylating Claviceps strains producing elymoclavine eliminate the free alkaloid into glycosides. The feedback inhibition of alkaloid synthesis by elymoclavine is then strongly reduced, helping to further improve elymoclavine yields. Elymoclavine can be liberated simply by invertase activity of baker's yeast.
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References
Dickerson AG (1972) A β-d-fructofuranosidase from Claviceps purpurea. Biochem J 129:263–272
Flieger M, Zelenkova NF, Sedmera P, Křen V, Novák J, Rylko V, Sajdl P, Řeháček Z (1989) Ergot alkaloid glycosides from saprophytic cultures of Claviceps. I. Elymoclavine fructosides. J Nat Prod 52:506–510
Flieger M, Křen V, Zelenkova NF, Sedmera P, Novák J, Sajdl P (1990) Ergot alkaloid glycosides from saprophytic cultures of Claviceps II. Chanoclavine-I fructosides. J Nat Prod in press
Floss HG, Günther H, Mothes U, Becker I (1967) Isolierung von Elymoclavin--O-β-d-fruktosid aus Kulturen des Mutterkornpilzes. Z Naturforsch 22b:399–402
Křen V, Pažoutová S, Rylko V, Sajdl P, Wurst M, Řehácĉek Z (1984) Extracellular metabolism of sucrose in a submerged culture of Claviceps purpurea: formation of monosaccharides and clavine alkaloids. Appl Environ Microbiol 48:826–829
Křen V, Ludvík J, Kofroňová J, Řeáček Z (1987) Physiological activity of immobilized cells of Claviceps fusiformis during long term semicontinuous cultivation. Appl Microbiol Biotechnol 26:219–226
Křen V, Břemek J, Flieger M, Kozová J, Malinka Z, Řeháček Z (1989) Bioconversion of agroclavine by free and immobilized Claviceps fusiformis cells. Enzyme Microb Technol 11:685–691
Perényi T, Udvardy-Nagy E, Novák EK (1968) Studies on sucrose breakdown by invertase-producing Claviceps strains. Acta Microbiol Sci Hung 15:17–28
Robbers JE, Cheng Li-jung, Anderson JA, Floss HG (1979) Synthesis of ergot alkaloids by protoplasts of Claviceps species. J Nat Prod 42:537–539
Spalla C (1973) Genetic problems of production of ergot alkaloids in saprophytic and parasitic conditions. In: Vaněk Z, Hoštálek Z, Cudlín J (eds) Genetics of industrial microorganisms. Academia, Prague, pp 393–402
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Křen, V., Flieger, M. & Sajdl, P. Glycosylation of ergot alkaloids by free and immobilized cells of Claviceps purpurea . Appl Microbiol Biotechnol 32, 645–650 (1990). https://doi.org/10.1007/BF00164733
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DOI: https://doi.org/10.1007/BF00164733