International Genetically Engineered Machine 2013 Competition - PowerPoint PPT Presentation

ITB_Indonesia International Genetically Engineered Machine 2013 Competition

Indonesia and risk of aflatoxin

Bad post harvesting and storage system can increase the growth of fungi such as Aspergillus sp % Commodity Contaminated by aflatoxin Egypt 11% 8% 3% Others 11% India Malaysia Nepal 48% 5% Nigeria 6% Turkey 8% Indonesia Farombi, et. al. (2006) Aflatoxin contamination of foods in developing ountries: Implications for hepatocellular carcinoma and chemopreventive strategies

Increasing susceptibility of liver cancer* *International Agency for Research of Cancer (IARC).

• Build a whole cell biosensor for aflatoxin B1 detection in foods

In monitoring environment status (Polutans, toxic compounds, etc) we need :  Accuracy  Rapidity Conventional Sensor VS Biosensor • Biosensor • Biomolecule • Whole cell

aflatoxin GFP

Bba_J23119 Signal peptide Bba_K1064004 CYP 3A4 B0030 Double terminator Illegal Restriction Site Bba_K1064000 Remove the illegal restriction site Remove the signal peptide Add a strong RBS Add a constitutive promotor

SDS PAGE analysis of CYP expression

Promotor sensitive To DNA damage Reporter Gene Double terminator pUV pSOS pSOS + GFP (Bba_K1064002) pUV + GFP (Bba_K1064003) Part submitted Part submitted

pUV pSOS Bright Field Microscope Fluorescence Microscope Bright Field Microscope Fluorescence Microscope Control (No UV) UV irradiation (10s)

pUV pSOS 62 60 58 56 Fold GFP 54 52 50 48 46 44 Bba_K1064002 (pSOS+GFP) Bba_K1064003 (pUV+GFP)

To find time for analyzing aflatoxin presence in food Goal with sensitivity threshold about 20 ppb* *) Based on Indonesia Food and Drug Regulation Simulate all phenomena! Enzymatic reaction Reporter Aflatoxin diffusion DNA damage and SOS response CYP synthesis

using Simbiology (computational biology toolbox from Mathworks Matlab) : minimum time needed for analysis 45 ppb 40 ppb GFP (mole) 35 ppb 30 ppb 25 ppb 20 ppb minimum observable [GFP] 15 ppb 10 ppb 5 ppb 0 ppb (s)

 Submitting part Bba_K1064002 and Bba_K1064003  Our construct works successfully as expected  Characterize part Bba_K1064002 and Bba_K1064003  Improvement on part BBa_J22106 and Bba_I765001

 Making prototype of our device  Adding another more visible chromopohore as reporter  Constructing the reactive sequence  amplifying the DNA damage signal  Adding buoyancy system  Make the device safer for user

 Indonesian Potential to Synthetic Biology synbio.itb.ac.id facebook.com/synbio.itb @synbio_itb

 Project • Survey : Indonesian people awareness about aflatoxin and the urgency of detector of it 32% correspondent need a biosensor

Dr. Sony ARI INDRA NUKE Dr. Maelita Biotechnology Microbiology Microbiology LILI DIMAS RIANDY Dr. Adi Bioengineering Chemical Informatic Engineering Engineering

Special Thanks To : • School of Life Sciences and Technology • School of Electrical and Informatics engineering • Faculty of Industrial Technology