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Application

There are many possible applications for our Sporobeads as it is possible to display all kinds of different proteins on their surface. To easily create any kind of Sporobead we designed a Sporovector, where you just have to insert your gene encoding your fusion protein of choice.

What are the potential advantages of using our B. subtilis spores as functional bioparticals?
They are

	easy and cheap to produce in large quantities (check out the link to this commertialoffer)
	very stable even under severe environmental conditions
	unable to proliferate and therefore most likely not treated as genetically modified organisms by the law
	part of human nutrition in terms of food supplements and generally regarded as safe [GRAS]

In the following paragraphs, we would like to present examplary ideas for possible applications of our Sporobeads.

Kumamolisin-Sporobeads: the solution for carefree enjoyment of everyday meals

World-wide, one out of 3350 people cannot eat meals that contain wheat products or other foods with traces of gluten. This disease is known as celiac disease. Kumamolisin is an enzyme that cleaves peptides and was produced by the iGEM-Team from the University of Washington last year. The substrate includes a specific sequence of amino acids which causes celiac disease in sensitive people when they consume food containing gluten. Our beads could carry Kumamolisin and offer a protected passage through the stomach, so that the enzyme can work properly where it is needed in the intestines. The GerminationSTOP we integrated in our spores would prevent outgrowth and ensure a correct dosage. This project is a pharmaceutical application and therefore would have to fulfill the legal requirements for pharmaceuticals. This includes several verification steps of non-toxicity and efficacy.

CPX-Sporobeads: rescuing marine life

The excessive use of disposable plastic and the lack of universal recycling programs has led to the pollution of the world's oceans. In the ocean, large pieces of Polystyrene litter are ground by sea currents into very small pieces, so called plastik plankton, that are consumed by fish, filter feeders, and other organisms living in the oceans. Such plastic uptake can lead to poisoning, sterility and death. The CPX-peptide can bind to Polystyrene and was generated by the iGEM team from MIT (2007). CPX-Sporobeads in huge filter boxes could be put into place to mechanically filter microscopic plastic particles, like polystyrene plankton, out of the water. To prevent the beads from being released into the sea and the plastic to be kept, the Sporobeads could be attached to membranes in the boxes. Then the spores would not only express CPX but also a membrane binding protein on their surface.

TALE-Sporobeads: easy and cheap detection of genetically modified organisms

Since 1990, green biotechnology releases many transgenic plants into the environment by selling genetically modified seeds. Thus organic farmers need to prove, that their products meet the requirements for organic crops. Usually they pay laboratories to attest the lack of contamination with genetically modified crops. New tools for sequence-specific DNA-binding of molecular biology, TAL effectors, combined with our Sporobeads could be an easy and cheap solution for organic agriculture. Farmers could use a kit with TALE-lacZ-Sporobeads to detect sequences specific for genetically modified crops themselves. As spores are stable and safe vehicles, they could be send by mail without any considerations. The kit would be suitable for use outside of laboraty. The protocol for this could work as follows:
DNA extracted from plants with solutions provided by the kit is immobilized and fixed on a nitrocellulose membrane. This membrane is then washed, incubated with Sporobeads in solution and washed again. With addition of a substrate, the lacZ of bound Sporobeads will catalyze a reaction so that a blue staining appears. If no such DNA is present, the spores will not bind and no blue color will appear.

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