SELEKSI DAN OPTIMASI SEL RAGI TRANSFORMAN DNA KINA (CINCHONA LEDGERIANA MOENS) DALAM MEMPRODUKSI SENYAWA ALKALOID KINA

• Main Author: Andriani (NIM 20695014), Robbiatun
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/5522
• Institution: Institut Teknologi Bandung

<b>Abstrak :</b><p align=\"justify\"><br /> An experiment on the selection and optimization of yeast cells transformed with Cinchona (Cinchona ledgeriana Moens) DNA to produce Cinchona alkaloid compound had been carried out in 3 steps procedure. The first step was to select the best transformant from 55 isolates which was divided into 4 master plates (A,B,C and D). All master plates were cultured on YEPDA (Yeast Extract Peptone Dextrose Agar) medium. One isolate was selected from each master plate, they were A4, B16, C39 and D50, all were able to produce alkaloid compound as showed by the positive reaction with Dragendorff reagent. The second step was to determine the optimal composition and concentration of the production media and the inoculum concentration for each selected transformed cells which produced alkaloid compound. Two liquid production media, i.e. YEPD and PD (Peptone Dextrose) were used at concentration of media 10%, 25%, 50% and 75% containing inducer 250 ppm L-tryptophan. The variation of inoculum concentration were 10%, 15% and 20%. The result showed that A4 and C39 isolates were able to produce an alkaloid compound. The optimal concentration for A4 were 50% PD as production medium and 10% inoculum while C39 in 50% YEPD and 20% inocuhum. The third step was to determine the alkaloid compound by Thin Layer Chromatography (TLC) methode according to ). The result showed the alkaloid compound was not quinine or qu inidine but it was assumed to be an anthraquinone compound or intermediate compound in Cinchona biosynthesis.