Esyanti, Rizkita Rachmi and Wulansari, Amilia and Dingse, Pandiangan
(2008)
THE INFLUENCE OF IMMOBILIZATION ON CELL GROWTH AND CATHARANTHINE PRODUCTION IN THE CELL AGREGATE CULTURE OF Catharanthus roseus (L.)G.Don.
Proceeding of 2nd International Conference on Mathematics and Natural Sciences (ICMNS) 2008.
pp. 172-177.
Abstract
Immobilization method is frequently used as one of the reliable techniques to increase in vitro secondary metabolite production. Research has been conducted to evaluate the influence of cell immobilization on cell growth and catharanthine production in C.roseus culture. Callus was first initiated in MS medium with 1,5 mg/L 2,4-D and 0,15 mg/L kinetin. The cell agregate derived from callus of Catharanthus roseus was cultured in MS medium with 2mg/L NAA and 0,2 mg/L kinetin, and subcultured every 10 days for 4 times before immobilization. Immobilization was conducted by using 2,5% Na-alginate. The cell growth and production pattern of catharanthine in the cells agregate were observed every 3 days for 27 days. Qualitative and quantitative analyses were conducted by using HPLC, eluted by using methanol : acetonitrile : diamoniumhydrogenphosphate (3:4:3), rate: 1ml/minute. The results showed that catharanthine was detected in the cells agregate and culture medium. The highest concentration of catharanthine in the control cell was detected on the 9th day of culture, i.e. 39,1843 μg/g, while in medium was obtained on the 12th day of culture, i.e. 5,6892 μg/L. Analysis on the immobilized cells showed that optimal content on the cells agregate was obtained on the 9th day (136,4230 μg/g), while in the medium was on the12th day of culture, i.e. 17,6306 μg/L. The average concentration of catharanthine in immobilized agregate cell culture increased 248 % compared to the control. The result also showed that catharanthine was still retained in the alginate matrix, and reached 3,6974 μg/L on the 18th day. Therefore further experiment is still needed to optimize the secretion of catharanthine into the medium.
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