Team:MSOE Milwaukee/Project


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Eucalyptol, our product of interest, is a monoterpenoid that can be used as an insecticide and an insect repellant. It is currently obtained in large quantities by fractional distillation of eucalyptus oil, which is obtained through the steam distillation of the leaves of selected Eucalytpus species, which are found primarily in China, but also in South Africa, Portugal, Spain, Brazil, Australia, and Chile. Eucalyptol comprises up to 90 percent of Eucalyptus oil, which has a variety of uses including flavoring in baked goods and beverages, preventing the separation problem with ethanol and petrol fuel blends, and it can even be used as a fuel. However, it is not currently used as a fuel because production costs are simply too high for this to be economically feasible. Eucalyptol has also been found to control airway mucus hypersecretion and asthma, inhibit cytokine production in cultured human lymphocytes, reduce inflammation and pain when applied topically, and kill leukemia cells.
Fuel Source Scent Industry Flavorings
It has been shown that Eucalyptol can be used as a fuel additive. It can be used in a 1:8 ratio with traditional gasoline so that the gasoline will last eight times longer! One man even added it to his Honda motorcycle with great results. He was able to drive around the city with the Eucalyptol and gasoline mixture in his tank, and no harm came to his engine. Eucalytpol is the major component of Eucalyptus oil, which is used for many fragrance applications. The most common uses in regard to fragrance are to create a fresh and clean aroma in perfumes, soaps, lotions, and detergents. Eucalyptol is used as a flavoring in many different products, including mouthwash and chewing gum. It is also used in low levels (0.002 percent) in items such as baked goods, meat products, and even beverages. It is also claimed to improve the flavor as an additive in cigarettes.
The three E. coli system can be separated into input and output. The input cells are responsible for breaking down the spent grains into sugars that the output cells can use. Spent grains are mainly composed of hemicellulose and cellulose. Our input cells contain genes xynC-A and xyloA that break down hemicellulose into xylose. However, hemicellulose is too big to enter the cell. The input cells are designed to tag the proteins from the two genes and secrete them out. Since there are two genes to tag, there are two different versions of the input cell. One contains just the xynC-A tag plasmid and pump and the other contains xyloA tag plasmid and pump. The input cells will create xylose in the media ready to be used by the output cells. The output cells have the same plasmids in them. The plasmids contain genes of the Mevalonate Pathway that are not present in E. coli. There are six in total with one coming from E. faecalis, three from S. pneumonia, one from Arabidopsis thaliana, and biobrick BBa_K849000. These organisms were chosen because they proved to be the most efficient genes in producing Isopentenyl-5-pyrophosphate. Our gene of interest, TPS-CIN, will use that product to make eucalyptol. The eucalyptol will stay in the cell or in the media and can be extracted.

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