Team:Austin UTexas/Demonstrate
| Species | Classification | Brand of Kombucha Isolated From |
|---|---|---|
| Staphylococcus warneri | Bacteria | GT’s Kombucha |
| Staphylococcus epidermidis | Bacteria | GT's Kombucha |
| Gluconobacter oxydans* | Bacteria | GT’s Kombucha |
| Lachancea fermentati* | Yeast | Buddha's Brew |
| Propionibacterium acnes | Bacteria | Buddha's Brew |
| Micrococcus luteus | Bacteria | Buddha's Brew |
| Bacillus pumilus | Bacteria | Buddha's Brew |
| Saccharomyces cerevisiae | Yeast | LIVE Soda Kombucha |
| Schizosaccharomyces pombe* | Yeast | LIVE Soda Kombucha |
(*Indicates a species that is considered vital to the production of kombucha)
pH Sensors
Possessing the ability to monitor the brewing process of kombucha without disturbing the microenvironment and using a very visible color reporter would allow for greater insights as to how the populations of organisms and pH may change due to competition amongst other bacteria and yeast in the beverage and SCOBY of the kombucha. The byproducts produced by the kombucha as it brews causes the tea to become more acidic, leading to our team searching for pH sensitive promoters, and for ways to implement these into kombucha.
Though an acidic sensor was what was required for our kombucha analysis, the identification of sensors in other areas of the pH spectrum were explored as well. Three sequences were identified, the CadC operon for the acidic range, CpxA-CpxR complex for the neutral range, and the P-atp2 promoter from the BioBrick Registry (BBa_K1675021) for the basic range. Each sequence was paired with a unique corresponding reporter sequence so that if each pH sensitive plasmid were in the same environment, the specific pH of the system could be seen. The reporters used were, BBa_E1010 for the CadC construct, BBa_K1033916 for the CpxA-CpxR complex, and BBa_K592009 for the P-atp2 promoter.
CadC
CpxA-CpxR
P-atp2
Next Steps and the GOX Sequences as Putative Promoters
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