Team:Bilkent UNAM Turkey/Project Description

Health concern and pollution created by TNT

TNT or 2,4,6-trinitrotoluene was produced in 1863 by Wilbrand, and since then used as the primary explosive power provider by military forces in the world. Military areas are the most frequent source of TNT contamination in the world.
TNT can penetrate through the skin and poses a health risk to human. The adverse effects of TNT were indicated for humans such as irritation of the skin, aplastic anemia, anemia, abnormal liver functions, spleen enlargement, cardiac failures, cataract or spermatozoa malformancy. TNT is reported to have high levels of carcinogenic and mutagenic properties. The dust of TNT causes severe respiratory disturbances and relapsing illnesses.
On the other hand, there is a high risk of contamination of food and water resources by TNT leakage from the contaminated areas. After hurtful usage of lands for military purposes, significant amounts of TNT or its derivatives accumulates on the lands. TNT and RDX has high tendency to travel into the water and its removal in natural ways lasts decades. TNT or TNT derivative RDX (cyclotrimethylenetrinitriamine) contaminated waters looks like pink colored which gives its name to this phenomenon as “pink water” or “red water”. It is very hard to clean them and cost very much.
Thus, removal of TNT by means of biological ways would be a great task to do

Our Motivation and Aim

Our motivation was to find biological ways to remove pollutants otherwise tightly bound to the soil and water. One of the best known pollutant of soil and water is TNT and cause great burden to the environment and human health. As of this year, there is not a comprehensive, economical and effective method to remove this pollutant from the water and soil.

Microalgae Phytoremediation

Both higher and lower plants (unicellular microalgae) are used for phytoremediation (removing pollutants by using plants) because of their great capability of absorbing and removing poisonous materials located in soil, water and air. Our aim is to use Chlamydomonas Reinhardtii to remove TNT and its derivatives from the soil and water. Our approach of phytoremediation includes the biotransformation of TNT and RDX to its less harmful derivatives.

Experimental Design

- Design and order of Synthetic nfsI according to C. reinhardtii codon bias table and biobrick standards.

- Construct algal protein expression vector (pRbc backbone and biobrick prefix and suffix).

- Built of pRbcnfsI for transfection of algae with pKS-aphVIII-lox plasmid for antibiotic resistance against
neomycin (cotransformation).

Expected Results

Microbial transformation of toxic TNT to non-toxic ADNT will be achieved using Chlamydomonas reinhardtii. This organism can be used to clean both contaminated soil and pink water since it is capable of living in potentially toxic environments.