Pengoptimuman kul tur amp alan sel eurycoma longifolia jack untuk penghasilan biojisim dan alkaloid
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Date
2004-08
Authors
Mahmud Siregar, Luthfi Aziz
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Abstract
Alkaloids, 9-hydroxycanthin-6-one and 9-methoxycanthin-6-one, could be detected
from Eurycoma longifolia Jack roots with gas chromatography-mass spectrometer
(GC-MS) and high performance liquid chromatography (HPLC). Biomass of E.
longifolia Jack callus and suspension cells were also found to contain 9-
hydroxycanthin-6-one and 9-methoxycanthin-6-one using the same methods. The
initiation of callus from the leave explants produced higher 9-hydroxycanthin-6-one
and 9-methoxycanthin-6-one compared with callus initiated from the petiole and stem
explants. Cell line was selected based on cell biomass and alkaloids (9-
hydroxycanthin-6-one and 9-methoxycanthin-6-one) contents in callus derived from
leaves of different plant. Cells derived from leaves of Eu9 plant were selected as elite
cell line although they contained moderate alkaloids but the cells could be initiated to
form fine cell suspension cultures and could be . easily maintained with repeated
subcultures compared with the other cell lines. The repeated subculture of Eu9 cell
suspension cultures for three years did not cause browning of culture, and did not
result in decrease of cell production and the content of alkaloids produced was stable.
The pH of culture medium fixed at 5.75 induced higher total alkaloids content.
Addition of Chitosan® as an elicitor into MS liquid medium + 2.69 11M NAA + 1.13
11M 2,4-D on the 131
h day of culture stimulated the increased production of 9-
methoxycanthin-6-one by 5.3 fold. While addition of 10 mg/L Orykta™ clay into MS
liquid medium+ 2.69 11M NAA + 1.13 11M 2,4-D resulted in two fold increased on
production of two alkaloids, 9-hydroxycanthin-6-one and 9-methoxycanthin-6-one.
Optimization studies of macronutrients, Fe-EDTA, micronutrients, sucrose
concentration and addition of cytokinin into MS medium for cell biomass and
production of alkaloids were carried out individually. The modified macronutrients,
Fe-EDTA, micronutrients and sucrose in MS medium based on optimum cell biomass
production produced MSB5 medium which was consisted of MS + 12.25 mM KN03,
0.575 mM MgS04AH20, 1.8 mM KH2P04, 0.11 mM Fe-EDTA, 0.11 mM
MnS04AH20 and 4 % sucrose. This MSBs medium was found to induce the highest
cell biomass and total 9-hydroxycanthin-6-one content in the cells within 14 days of
culture under light intensity of 1525 lux. The modification of macronutrients, FeEDT
A, micronutrients and sucrose concentration in MS medium with the addition 2.32
11M kinetin based on the production of the highest amount of the two alkaloids content
produced MSF medium. MSF medium consisted of MS + 21.5 mM ~N03 , 14.25
mM KN03, 7.1 mM CaCh.2H20, 2.5 mM MgS04.4H20, 1.45 mM KH2P04, 0.233
mM Fe-EDTA, 0.215 mM MnS04AH20, without CuS04.SH20, 4% sucrose+ 2.32
11M kinetin stimulated low cell biomass production but produced the highest
production of the total 9-methoxycanthin-6-one in the cells of E. longifolia Jack. To
obtain high cell biomass of E. longifolia Jack with high production of 9-
methoxycanthin-6-one, the cells should be cultured using two stage culture system.
The cells were cultured in MSBs liquid medium in the first stage then followed by
culturing the cells in MSF liquid medium in the second stage.
Description
Keywords
Ampaian sel eurycoma longifolia , Penghasilan biojisim alkaloid