The Enhancment of Andrographolide Content from Andrographis paniculata (Burm.f.) Wallich ex Ness Cell Culture by Using Immobilized Cell in Bioreactor

• Main Author: Ramdaningsih Kadar, Vidia
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/34861
• Institution: Institut Teknologi Bandung

Andrographis paniculata is known to contain andrographolide, a secondary metabolite which shows pharmacology effects such as hepatoprotective, antiviral and anticancer. Previous reports indicated that the content of andrographolide produce in tissue culture system was lower than that produced in the intact plant. Thus, in order to increase the amount of andrographolide in tissue culture system, cell immobilization was adopted. A bubble column bioreactor was used to scale-up cell biomass as well as andrographolide production. The aims of this research were to increase the amount of andrographolide by using cell immobilization, and to scale-up cell biomass by using bioreactor. Cell culture was initiated by transfering a friable callus from semisolid medium into liquid Murashige & Skoog medium with the addition of 3.0% sucrose, 0.1 ?M BAP (6-benzylaminopurine) and 0.5 ?M 2,4-D (2,4-dichlorophenoxyacetic acid) in an erlenmeyer flasks which were agitated at 120 rpm (rad per minute) at room temperature. After ten days, a part of cell culture was immobilized in 2.5% alginate to form beads of cells in matrix of alginate. To scale-up cell biomass, 35 g fresh weight of cells was transferred into a bubble column bioreactor with 320 mL/min aeration at room temperature. Immobilized cells were also transferred into bioreactor to scale-up production of andrograpollide. The growth of cells in the erlenmeyer and bioreactor were observed based on their dry weight (DW) as well as the immobilized cell growth. Andrographolide content in the samples of cell culture and immobilized cell were analyzed by using HPLC (High Performace Liquid Chromatography) equipped with UV detector at 254 nm and ODS-C18 column. Elution was performed by a mixture of methanol-water (7:3) at the flow rate of 0.5 mL/min. The observation of growth indicated that the growth of immobilized cell was slower than that of cell culture in the erlenmeyer. The highest amount of andrographolide produced in erlenmeyer was observed as much as 5.57 mg/g DW and 9.69 mg/g DW in the cell culture and immobilized cell, respectively. In the bioreactor, the content of andrographolide was 1.51 mg/g DW and 4.23 mg/g DW in the cell culture and immobilized cell, respectively. This results indicated that cell immobilization could enhance the content of andrographolide at two times and three times higher in erlenmeyer and bioreactor, respectively. A trace amount of andrographolide was also detected in the medium cell culture and vi medium immobilized cell as much as 0.00083 mg/mL and 0.004 mg/mL, respectively.