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Journal > Jurnal Bahan Alam Indonesia > PENINGKATAN PRODUKSI ALKALOID INDOL KANTINA-6-ON DI DALAM KULTUR SUSPENSI Brucea javanica (L.) Merr.

 

Jurnal Bahan Alam Indonesia
Vol 1, No 1 (2002)
PENINGKATAN PRODUKSI ALKALOID INDOL KANTINA-6-ON DI DALAM KULTUR SUSPENSI Brucea javanica (L.) Merr.
Alam, Germini ( Fakultas Farmasi Universitas Gadjah Mada)
Wiryowidagdo, Sumali ( Jurusan Farmasi FMIPA Universitas Indonesia)
Soegihardjo, C J ( Fakultas Farmasi Universitas Gadjah Mada)
Sudarsono, . ( Fakultas Farmasi Universitas Gadjah Mada)
Gobel, R B ( Jurusan Biologi FMIPA Universitas Hasanuddin)
Article Info   ABSTRACT
Published date:
01 May 2012
 
In an attempt to increase the productivity of the pharmaceutical compound canthin-6-one from cell suspension culture of Brucea javanica, a high cell density culture and improved culture conditions have been investigated.  The  effect  of  frequency  of  sub-culturing,  concentration  of  2,4-D,  addition  of  tryptophan  as  a stimulant, and culture stage of growth on the production of canthin-6-one, was of particular interest in this study. The highest yield of canthin-6-one, 26.37 mg g-1 dry cell, was produced intra cellular in 50 g L-1 cell mass.  The  established  cell  suspension  were  harvested  at  the  age  of  60  days  and  placed  into  MS  medium containing 0.2 mg L-1 2,4-D and 10 ml L-1 tryptophan for 33 days, approximately when the cultures were at their late stationary phase of growth. Addition of 20 mg L-1 tryptophan did not show a significant difference in canthin-6-one production (p>0.05). Higher than 20 mg L-1 or lower than 10 mg L-1 of tryptophan showed adverse  effect  on  the  concentration of canthin-6-one alkaloid (p>0.01). Qualitative analysis of chloroform-extract  of  dried  cells  from  suspension  cultures  revealed  eight  spots  on  TLC.  The  identity  of  canthin-6-one was  established  by  comparison  of  its  Rf  value  (Rf  =  0.37),  its  color  reaction  with  Dragendorf  reagent  and ceric  ammonium  sulphate,  and  its  behavior  in  UV  with  an authentic sample. Quantitation of canthin-6-one alkaloid using TLC-densitometry scanner in response to different concentration of tryptophan as a precursor showed  a  concentration-dependent  response.  The  production  of  canthin-6-one  at  26.72  mg  g-1 dried  cell weight  in  this  study,  establishes  a  methodology  of  an  improved  procedure  compared  to  those  previously reported.
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