1. 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

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

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

4. 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.

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

6. 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

7. 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

8. Entrapment in calcium alginate Immobilized Catharanthus roseus cells

9. Indole alkaloids biosynthesis

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

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

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

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

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

15. 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

16. 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.

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

18. Thank you for attention!
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