Team:LMU-Munich/Application

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Revision as of 18:44, 25 October 2012

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

They are:

	Very stable even under severe environmental conditions
	Easy and cheap to produce in large quantities (check out the link to this commercial [http://www.alibaba.com/product-free/118791994/Bacillus_subtilis_spores_HU58_100_pure.html offer])
	Part of human nutrition in terms of [http://en.wikipedia.org/wiki/Bacillus_subtilis_R0179 food supplements] and [http://www.fda.gov/Food/FoodIngredientsPackaging/GenerallyRecognizedasSafeGRAS/default.htm generally regarded as safe]
Moreover, our Sporobeads are:
	Unable to proliferate and therefore most likely not treated as genetically modified organisms by the law

Our Sporobeads offer a wide variety of applications within the laboratory, and around the world. Sporobeads can be used for filtration and protein screening.

Filtration

Further information for the first possible application for our Sporobeads.

Filtration

Our Sporobeads can express proteins on their outer coats to bind specific molecular targets. Such targets include heavy metals, toxins, and plastic.

One example of such a filtering protein could be a CPX-Sporobead. [http://partsregistry.org/wiki/index.php/Part:BBa_I728500 CPX] is a peptide developed by the 2007 MIT iGEM team, which is extremely hydrophilic, and thus capable of binding microparticles of polystyrene from water. The excessive use of disposable plastic and the lack of universal recycling programs has led to the [http://www.ncbi.nlm.nih.gov/pubmed/22610295 pollution of the world's oceans]. In the ocean, large pieces of plastic litter are ground by sea currents and degraded by UV radiation into microscopic pieces, so called "plastic plankton," which is consumed by fish, filter feeders, and other marine organisms. Such plastic uptake can lead to poisoning, sterility and death.

CPX-Sporobeads in huge filter boxes could be put into place to mechanically filter microscopic plastic particles out of the water. Such specific filtration would be superior to blanket filtration systems, which also remove living phytoplankton important to ocean ecosystems. To prevent the beads from being released into the sea and to ensure the plastic be removed from the water, the Sporobeads could be attached to membranes in the filter boxes. Then the Sporobeads would need to not only display CPX but also a membrane binding protein on their surface.

Protein Screening

Further information for the possible application of protein screening.

Protein Screening

Designer protein molecules, which bind their desired targets specifically, have numerous uses. Designer proteins are frequently created by constructing and screening libraries of mutated protein variants. Proteins with desired properties are selected for in this method. Despite the success of the method, it is labor intensive and limited in throughput. Individual mutated proteins must be tracked throughout the entire screening process. Our Sporobeads would eliminate the need to track protein mutants, allowing researchers to screen huge quantities of mutated proteins, only identifying and sequencing these proteins after successful screening. Figures 1 and 2 offer schematics of the process of using Sporobeads for protein screening for tests and affinities.

Fig. 1: The simple protein-screening process in our Sporobeads.

Fig. 2: The simple protein-screening process in our Sporobeads using affinity binding as a requisite.

Further Applications

Further information for the possible application of protein screening.

Further Applications

Ultimately, the ability to express virtually any protein of interest shows great potential for laboratory work. Our Sporobeads could additionally express TAL effectors, for the binding of sequence-specific DNA stretches. This could allow simple GMO detection of food crops. Besides simply expressing proteins capable of binding elements of interest, our Sporobeads could express proteins which have enzymatic activity. The 2011 University of Washington iGEM team developed an enzyme, Kumamolisin, which cleaves peptides. The specific substrate of this enzyme is the specific amino acid sequence which causes reactions in individuals with Celiac disease. Such a cleaving enzyme could be used to eliminate irritants from gluten-containing food products.

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 them, we designed a Sporovector in which you just have to insert the gene encoding your fusion protein of choice. Since there are so many possible applications, we illustrate three examplary ideas for future Sporobeads in the following section:

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