Team:Dundee/Project

Algae are photosynthetic organisms which live in aquatic environments. Cyanobacteria are also known as blue-green algae and they are prokaryotes that are responsible for the majority of photosynthesis on Earth. During the hot and sunny summer months we experience a phenomenon known as an algal bloom. This is a spectacular increase in the size of the algal population in a water body as the algae take advantage of the seasonal spike in light and warmth. With increased amounts of nutrients leaching into water due to agriculture, blooms are becoming more and more common. Aside from affecting the environment they occur in, algae can also be dangerous for humans as many species produce toxins. We have decided to focus on one hepatotoxin called microcystin, a cyclic non-ribosomal peptide that binds covalently and irreversibly to protein phosphatases in mammalian bodies, inactivating them.

The study of biochemical processes in human cells is one of the most heavily-researched scientific fields. However, it is not often that scientists exploit the biochemical potential that our bodies offer in order to create new technologies for environmental remediation. We have decided to exploit the human protein phosphatase 1 (PP1)–microcystin interaction to create a bacterium that will sequester microcystin and prevent its toxic action.

We engineered the chassis organism Eschericha coli to export human protein phosphatase 1 (PP1) to its periplasmic compartment. By doing this we have created a biological mop for microcystin that we call the ToxiMop.

Sensor

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Detector

One of our aims is to make a detection system in Bacillus subtilis where spores germinate in response to the presence of microcystin. This uses a modified version of the PrkC receptor (which usually acts as a sensor for the germination of other spores in the surrounding area), substituting PP1 in place of the normal sensor domains. Our idea is that when microcystin binds to the PP1 of the PrkC receptor, this will trigger downstream intracellular effects, resulting in the germination of dormant spores. The awoken bacteria will then produce PP1 (ToxiMop idea) to rid the water of microcystin.

Toxi-Mop

ToxiMop uses the human protein PP1 expressed on the surface of bacteria to act as a biological mop for microcystin. By covalently binding to protein phosphatase 1 (PP1), microcystin acts as an inhibitor of PP1, a protein which is integral to almost all major signalling pathways in human cells. We are using cloning techniques to genetically engineer B. subtilis and E. coli to express PP1 on the cell surface so that bacteria can inhibit microcystin in algal blooms, therefore reducing harm to freshwater ecosystems.

Toxi-Tweet

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