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From 2009.igem.org

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PROJECT - Microbial Fuel Cell: The goal of this research is the manipulation of E.Coli; granting them the ability to release electrons in an aerobic enviroment. This project utilizes geobacter's cytocromes, which makes it's extracellular electron transfer possible. The cytocromes OmcB,OmcE,OmcS, and MacA are our priority since they are the major electron smugglers out of the cell. By isolating and placing these cytocromes, some of which are naturally occur in E.Coli,into bricks we hope to make the electical application possible in other bacterial species.

File:Echain.JPG Extracellular Electron Transport Chain

Overall project

Optimization of electron shuffle to external surfaces such as anodes is a primary goal. Geobacter sulfurreducens happens to be our model bacteria due to its ability in nature to efficiently export electrons extracelluarly. E. coli can be the chassis for this experiment due to its genome already containing some key proteins in our preferred pathway. The proteins, such as extracellular pilin, MacA, and many other cytochromes, which E. coli does not have will be isolated from Geobacter sulfurreducens and introduced into E. coli to formulate the most optimal pathway for generating electronmotive force in a microbial fuel cell apparatus.

Some problems will be faced concerning plasmid engineering and the simple fact that Geobacter is anaerobic and E. coli is aerobic. Also, the role of Geobacter's pili in extracellular electron transfer is not clearly understood and could create a significant problem since e. coli does not have such pili. As a team, we will push in the right direction harder than an emf on the internal resistivity of a toroid. Many diverse team members will work in concert utilizing Missouri S&T’s dominating Electrical, Chemical, and Biological Engineering undergraduates along with Biological Science masterminds.

Project Details

Procedure: Some of the techniques that have been used thus far in pursuing this project have been: PCR amplification of the MacA and other cytocromes using PCR primers that added standard prefix and suffix sequences Cloning the cytocrome PCR products into a commercial cloning vector Digesting the cytocrome plasmids to obtain the promoter genes of interest Separating and isolating the enzyme digest fragments through gel electrophoresis Ligating the individual cytochrome promoters together Transforming bacterial cells with the recombined cytochrome plasmids

Possible Applications: The application of thie eletron trasfer is limited only by the imagination. The cytochrom cocktail could be used as a type of signal in a bacteria. It would produce small currents and be utalized in a similar fasion to the applications of flourescent proteins. Small electronics could uses living power sources (with future technological advances). Medical arenas could benefit by using modified bacteria in areas of the body such as stomach and intestines and developing the bacteria as a long term mobile monitor.

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