Molchan O.V., Romashko S.N. and Yurin V.M. The regulation of pharmaceutically active indole alkaloids biosynthesis in immobilized plant cells of Apocynaceae family. Belarussian State University, Minsk, Belarus Vinca minor

Catharanthus roseus G. Don Catharanthus roseus G. Don vinblastine are used in the treatment of Hodgkin disease, leukaemia are expensive (eur. 1.000.000-3.500.000 per kg) vincristine ajmalicine

Vinca minor L. are marketed as cerebral vasodilators vincamine, its derivatives, sum of vinca alkaloids are marketed as cerebral vasodilators

Plant cell and tissues cultures Callus and suspension cultures of Catharanthus roseus and Vinca minor can produce indole alkaloids initiation initiation callus tissue suspension culture But the yield of vincristine, vinblastine, ajmalicine and vincamine has been lower than that from the plant.

IMMOBILIZATION OF PLANT CELLS Alginate Polyacrylamide Agar and agarose Polyuretane Adsorption Covalent linkage Entrapment

Plant cell and tissues cultures Low productivity of the target substance Slow growth of plant cells Instability of cells in culture Difficulties in large-scale cultivation

A number of advantages of immobilized plant cells It encourages product release To affect secondary metabolite production It gives good cell-to-cell contact It protect the cells from shear stress

Entrapment in calcium alginate Immobilized Catharanthus roseus cells

Indole alkaloids biosynthesis

Tryptophan decarboxylase activity \freely suspended and calcium alginate-entrapped cells of Catharanthus roseus\

Tryptamine content \freely suspended and calcium alginate-entrapped cells of Catharanthus roseus\

Tryptamine content \incubation medium of freely suspended and calcium alginate-entrapped cells of Catharanthus roseus\

TDC activity and tryptamine release \freely suspended and calcium alginate-entrapped cells of Catharanthus roseus\

Alkaloids production \freely suspended and calcium alginate-entrapped protoplasts of Catharanthus roseus\

Vincamine production \incubation medium of freely suspended and calcium alginate-entrapped cells of Vinca minor\ 1 – susp 1 w 2 – imm 1 w 3 – susp 2 w 4 – imm 2 w 5 – susp 3 w 6 – imm 3 w 7 – leaf 8 - vincamine vincamine 1 2 3 4 5 6 7 8

Catharanthus roseus (L) G. Don, Vinca minor L are pharmaceutically important plants - a rich source of anti-cancer and anti-hypertensive indole alkaloids. Advantages the products (indole alkaloids) could be isolated from the incubation medium, rather then extracted from the biomass; the product could be removed continuously or semicontinuously from the incubation medium. the biomass does not destroy; the productive capacity of the system would not depend on the cellular accumulative capacity, but on the biosynthetic rate and rate release to the medium; it would be advantages since could be obtain high–producing immobilized cell cultures of Apocynaceae plant excreting indole alkaloids to the incubation medium.

REFERENCES Allen I. Laskin Enzymes and Immobilized Cells in Biotechnology. 1985. The Benjamin/Cummings Publishing Company, Inc. J.W. Pollard, J.M. Walker Methods in Molecular Biology, V. 6 Plant Cell and Tissue Culture. 1990. Humana Press. Clifton, New Jersey

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