PROJECT

Natural promoters sequence sensible to heavy metal in general have an unique bidirectional molecular organization. In the case of mercury, this complex structure is regulated by MerR repressor protein which binds to mer operator overlapped to its bidirectional promoter, having a weak expression. To apply this impressive mechanism in SynBio engineering systems is important to build new strong and well regulated genetic circuits to deeply understand its molecular interactions and to increase and improve its expression. We constructed the first library of new promoters regulated by MerR! This collection is well validated with RFP measurement and the most promising ones characterized with mercury clhoride. The best one passed the fermentation bioreactor test!

We adopted a strategy that goes beyond of what was presented before in the competition. We aim to enhance Mer Operon expression, improving previous synthetic genetic circuits and allying them with the construction of a bioreactor for water contaminated by mercury. With our genetic construction we achieved more than 97% of mercury bioremediation in 10 hours and then we made available diverse novel Biobricks for genetic constructions related to bioremediation.

Phytochelatin are proteins which have strong affinity to metals due its molecular composition with cystein aminoacids. For heavy metal bioremediation, was shown that recombinants with cellsurface display enriched with chelant proteins have better performance in adsortion of metalic ions. Nowadays there are several strategies to anchor proteins in membranes. In this project we used the most abundant cell membrane protein in E. coli, the outer membrane A (OmpA) fused to a synthetic phytochelatin codon optimized by our team in order to bio-accumulate mercury. We had awesome results and then we made this new synthetic phytochelatin designed by our team available!

We built the first (real) bioreactor to treat mercury contaminated water using our chassis carrying synthetic genetic circuits we developed! We had awesome experience working with fermentation experts and see our system working. We achieved the ultimate proof of concept and it was the first step to have this synbio system working out there in the real world.

In a selective environment, organisms adapt in order to survive. With rivers pollution by mercury, near and inside Manaus, wild bacteria have acquired resistance to this metal. We isolated and identified mercury resistent bacteria from our rivers. Identifying an microorganism never isolated before in Amazonia and sequenced the first Amazon genome in this competition!