Catharanthus roseus cells

Decendit, A., Liu, D., Quelhazi, L., Doireau, P., Merillon, J. M. and Rideau, M. 1992. Cytokinin-enhanced accumulation of indole alkaloids in Catharanthus roseus cell cultures. The factors affecting the cytokinin response. Plant Cell Reports, 11 400 03. 

Nef, C., Rio, B. and Chrestin, H. 1991. Induction of catharanthine synthesis and stimulation of major indole alkaloid production by Catharanthus roseus cells under non-growth-altering treatment with Pythium vexans extracts. Plant Cell Reports, 10 26-29. 

Courtois, D. and Guern, J. 1980. Temperature response of Catharanthus roseus cells cultivated in liquid medium. Plant Science Letters, 17 473 82. 

Verpoorte R, van der Heijden R and Moreno PRH (1997) Biosynthesis of terpenoid indole alkaloids in Catharanthus roseus cells. The Alkaloids, Chemistry and Pharmacology (ed Cordell GA) Vol 49. Academic, San Diego, pp 221-299. 

Cathenamine (100) has been identified as an early intermediate in terpenoid indole alkaloid biosynthesis (cf. Vol. 8, p. 27). It has also been isolated from Guettarda eximia. Another alkaloid, 4,21-dehydrogeissoschizine (99), has now been isolated from this plant it is readily converted into (100) in alkaline solution.29 On incubation with an enzyme preparation from Catharanthus roseus cell cultures in the presence of NADPH at pH 7, (99) was converted into ajmalicine (102), 19-ep/-ajmalicine (103), and tetrahydroalstonine (104), which are the normal products with this enzyme preparation. In the absence of NADPH, cathenamine (100) accumulated.30 The reaction to give (100) proceeded linearly with time, and was dependent on the concentration of protein and substrate. No conversion occurred in the absence of enzyme. 

Fig. 1 First example of a complete biosynthetic pathway to an alkaloid (ajmalicine = raubasine) based on enzymatic reactions elucidated by isolation of the single enzymes from Catharanthus roseus cell suspension cultures...

The integration of elicitation, in situ product adsorption with XAD-7, and the immobilization of Catharanthus roseus cells lead to an increase in productivity and a significant increase in extracellular ajmalicine production [5]. The integration of in situ product separation by two-phase culture and immobilized plant cells could be feasible for continuous production in immobilized plant cell bioreactors requiring the repeated use of cells. 

LUIJENDIJK, T.J.C., STEVENS, L.H., VERPOORTE, R., Purification and characterization of strictosidine (3-D-glucosidase from Catharanthus roseus cell suspension cultures. Plant Physiol. Biochem., 1998,36,419-425. 

GANTET, P IMBAULT, N THIERSAULT, M., DOIREAU, P Necessity of a functional octadecanoic pathway for indole alkaloid synthesis by Catharanthus roseus cell suspensions cultured in an auxin-starved medium. Plant Cell Physiol., 1998,39,220-225. 

The enzyme STR1 that was first characterized in Catharanthus roseus cell suspension cultures produces the central indole alkaloid intermediate H-3-a-(S)-strictosidine from tryptamine and secologanin (Fig. 8.9). It is well known that strictosidine represents the central intermediate precursor for several thousand indole alkaloids found in Nature. STR1 was the first gene to be cloned from R serpentina that involved a committed step in alkaloid biosynthesis.31 This was soon followed by the identification and isolation of an STR clone from Catharanthus roseus32 whose sequence was 80 % identical to the same gene from R serpentina.31... 

CONTIN, A., VAN DER HEIJDEN, R., LAFEBERE, A.W., VERPOORTE R., The iridoid glucoside secologanin is derived from the novel triosephosphate pathway/pyruvate pathway in Catharanthus roseus cell culture. FEBS Lett., 1998,434,413-416. 

Grabowski, L., Heim, S., and Wagner, K.G., 1991, Rapid changes in the enzyme activities and metabolites of the phosphatidylinositol-cycle upon induction by growth substrates of auxin-starved suspension cultured Catharanthus roseus cells. Plant Sci. 75 33-38. 

Endo, T., Goodbody, A., Vukovic, J. and Misawa, M. (1986) Enzymes from Catharanthus roseus cell suspension cultures that couple vindoUne and catharanthine to form 3, 4 -anhydrovinblastine. Phytochemistry, 27,2147-9. 

Rischer, H., Oresic, M., Seppanen-Laakso, T., Katajamaa, M., Lammertyn, R, Ardiles-Diaz, W., von Montagu, M.C.E., Inze, D., Oksman-Caldentey, K.-M. and Goosens, A. (2006) Gene-to-metabolite networks for terpenoid indole alkaloid biosynthesis in Catharanthus roseus cells. Proc. Natl. Acad. Sci. USA, 103,5614—9. 

Initially, the BR bios mthetic pathway was elucidated in Catharanthus roseus cell cultures by analysing the conversion products and intermediates (Fujioka et al, 1997). More recently, the biosynthesis of BRs has mainly been studied in Arabidopsis thaliam. Many of the genes encoding BR biosynthetic enzymes have been cloned using BR biosynthesis mutants of Arabidopsis thaliana, pea, tomato and rice. These mutants are BR deficient and revert to a wild-type phenotype following treatment with exogenous BRs. 

The terpenoid indole alkaloids have a variety of chemical structures and a wealth of biologic activities (Fig. 2a) (59, 60). Terpenoid indole alkaloids are used as anticancer, antimalarial, and antiarrhythmic agents. Although many biosynthetic genes from this pathway remain unidentified, recent studies have correlated terpenoid indole alkaloid production with the transcript profiles of Catharanthus roseus cell cultures (61). 

El-Sayed M, Choi YH, Frederich M, Roytrakul S, Verpoorte R. Alkaloid accumulation in Catharanthus roseus cell suspension cultures fed with stemmadenine. Biotech. Lett. 2004 26 793-798. Heinstein P, Hofle G, Stockigt J. Involvement of cathenamine in the formation of N-analogues of indole akaloids. Planta Med. 1979 37 349-357. 

A two-litre airlift type fermenter was also applied to the scaled-up culturing in the same conditions as the RDF. However, the shoot formation frequency was below 30%. Tanaka [28] reported that Catharanthus roseus cells were damaged by hydrodynamic stress when cultured in a jar fermenter but not when cultured in a RDF. The low... 

Deus-Neumann, B. and Zenk, M. H., Instability of indole alkaloid production in Catharanthus roseus cells in suspension cultures, Planta Med., 50 427 31 (1984)... 

The biotransformation of 15, 20 -anhydrovinblastine (207) in Catharanthus roseus cell suspension cultures (cell line 916) results in the formation of leurosine (208) (31%) and... 

Isochorismate synthase, that might be involved in the biosynthesis of 2,3-DHBA, has recently been purified from Catharanthus roseus cell-suspension cultures and subsequently its gene was cloned (L. van Tegelen, P. Moreno, A. Croes, G. Wullems and R. Verpoorte, submitted for publication). Two isoforms of the enzyme were purified and characterized. Both have an apparent molecular mass of 65 kD. The Km values for chorismic acid are 558 pM and 319 p.M for isoform I and II respectively. The enzymes are not inhibited by aromatic amino acids and require Mg for enzyme activity. The isolated cDNA encodes a protein of 64 kD with a A-terminal chloroplast targeting signal. The deduced amino acid sequence shares homology with bacterial isochorismate synthases, and also with anthranilate synthases, another chorismate utilizing enzyme. 

BIOSYNTHESIS OF TERPENOID INDOLE ALKALOIDS IN Catharanthus roseus CELLS... 

Effect of Different Treatments on Alkaloid Accumulation in Catharanthus roseus Cell Suspension Cultures"... 

Fig. 19. Effect of various signals on secondary metabolism in Catharanthus roseus cells.

P. R. H. Moreno, Influence of stress factors on the secondary metabolism in suspension cultured Catharanthus roseus cells. Ph.D. Thesis, Leiden University, 1994. 

Fig. 4. Growth of Catharanthus roseus cells in batch culture. (A) Concentrations of glucose (squares) and biomass (circles), (B) oxygen consumption, and (C) carbon dioxide production are given as a function of culture time. Vertical bars represent the standard deviation of the mean. Solid curves represent the model predictions.

Fig. 5. Changes in the amounts of intracellular carbohydrates in Catharanthus roseus cells during growth in batch culture.

Influence of Plant Growth Regulators on Alkaloid Production in Catharanthus roseus Cell Cultures... 

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