For our project, we are modifying TOP10 E. coli (MC1061 derivative), Gluconoacetobacter sp. (Komagataeibacter hansenii), and Gluconobacter oxydans. We are also working with (but not modifying) Lachancea fermentati and Saccharomyces cerevisiae. According to DSMZ, a German collection of information on microorganisms and cell cultures, all of our organisms are categorized as Risk Group 1 (no risk), with the exception of Lachancea fermentati, which has not been categorized because it is a multicellular fungus. Our TOP10 E. coli was acquired from the Barrick lab; Gluconoacetobacter sp. and Gluconobacter oxydans were ordered from ATCC (American Type Culture Collection) or isolated from kombucha; and Lachancea fermentati and Saccharomyces cerevisiae were isolated from kombucha. All of our strains except TOP10 E. coli are being used in recreating kombucha from scratch, as they are all naturally found in the beverage. The TOP10 E. coli are being used to develop our pH promoters which will then be inserted into the Gluconoacetobacter sp. and Gluconobacter oxydans, our chassis.
When working in the lab, everyone wears gloves to ensure protection from any laboratory chemicals and to minimize contact with living organisms. Furthermore, all members of the team know where safety equipment is in case of emergency. Waste is disposed of in containers appropriate to the type of waste. Additionally, to minimize the risk that somebody might think that our kombucha is okay to drink (even though eating and drinking are prohibited in the lab space), it is clearly labelled with identification and with a warning not to drink. Also, because our product will eventually be produced in factories for consumers to drink, other risks that our project might pose would be that the genetically modified organisms act in an unforeseen way or that a consumer might have an allergic reaction to one of the inserted genes. To reduce these risks, we will diligently observe our genetically modified organisms to ensure that they perform only the actions that we desire. If we observe the organisms doing something unexpected, we will troubleshoot according to the specific behavior they are displaying. As for people having a bad reaction to our product, we will include on our kombucha's label where the genes come from to reduce the likelihood that somebody with an allergy will come into contact with something that they shouldn't.