What is your chassis organism?

E. coli K-12 JM109, DH5α, BL21

Do you plan to experiment with any other organisms, besides your chassis?

In our project, we will make a food preservation box with antibacterial fragrances such as geraniol and farnesol that are produced by engineered E. coli. To assess an effect of antibacterial fragrances against microorganisms, we utilize and Bacillus subtilis var. natto as well as E. coli.

What risks does your project pose at the laboratory stage? What actions are you taking to reduce those risks?

The marA activating AcrAB-TolC efflux pump, also efflux antibiotics from engineered E. coli, resulting more resistance of the engineered E. coli to antibiotics. Because the host E. coli has no toxic to human, there may be no risk to human safety. All engineered E. coli have been sterilized by autoclave sterilization. We use hexane to dilute geraniol and farnesol and use decane to collect antibacterial fragrances from E. coli culture. Hexane and decane are flammable. These solvents are not used beside the fire. We have worn rubber gloves and have sterilized all wastes to reduce the risks, including safety level 1 procedures.

What risks might your project pose, if it were fully developed into a real product that real people could use? What future work might you do to reduce those risks?

The food preservation box have the risk that the engineered organisms grow in food. We must need to separate them from food area by packing or separating with membrane filters that the engineered organisms cannot pass through. Only fragrances should be dispersed into food area. This approach will decrease the risk of food contamination by engineered organisms. In our project, we have used E. coli JM109 strain, a derivative of E. coli K-12 strain encoding auxotrophy, which cannot grow in natural field. The engineered organisms will be used under supplication of required nutrition, so they cannot grow outside the box.