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Revision as of 21:23, 16 October 2016
Kombucha is a fermented tea that contains a symbiotic community which is characterized by the relationship between ethanol-producing yeast and bacteria. These different species of bacteria create bacterial cellulose and convert the ethanol produced from yeast, into acetic acid. Due to this unique microbiome, many claims have been made regarding the health benefits that come from drinking this beverage. However, none of these claims have been proven scientifically. Even so, many are quick to to jump on the bandwagon and want to try to create a profit from the rising popularity of kombucha. The Kombucha Brewers International, a non-profit trade association, has reported a growth of 50% per year in the kombucha industry with no signs of slowing down in the near future.1 Because of the growing popularity of kombucha and the fact that it has such a diverse community of microbes, we believe that our research can not only add to the field of synthetic biology, but help grow the industry of genetically modified foods. Our goal is to create a designer beverage with added benefits that come from the genetic modification of the microbiome inside. The following outlines the steps taken to achieve our goal.
1. Isolate and identify different microbes in kombucha through various growth mediums and antibiotics, while using 16s sequencing to reveal the identities of the microorganisms.
- So far, we have identified the yeasts Lachancea fermentati and Schizosaccharomyces pombe and the bacteria Gluconobacter oxydans.
2. Prove that genetic engineering is possible with the bacteria in kombucha by using conjugation to transfer a plasmid with a gene that produces GFP (Green Fluorescent Protein)
- We are still attempting conjugation, however, there has yet to be a successful attempt in Gluconobacter oxydans or Gluconacetobacter hansenii ( a close relative of Ga. xylinus).
3. Confirm successful conjugation by utilizing 16s sequencing to reveal the identities of the potential transconjugants.
- We have only sequenced E. coli, which means conjugation has not been successful.
4. Design a construct(s) in bacteria endogenous to kombucha that adds a beneficial aspect to the drink.
- We are currently in the process of designing a construct that produces Brazzein, a sweet tasting protein that can serve as an artificial sweetener. Another construct would include a part that increases the efficiency at which G. oxydans converts ethanol into acetic acid in order to decrease the ABV of the beverage.
5. Recapitulate create kombucha from scratch by adding specific strains of bacteria and yeast, including the transconjugants that contain our construct(s).
- We have successfully recapitulated kombucha with a mixture of our strains that has been isolated from store. However, due to the fact that there has not been a successful conjugation, there has not been a recapitulation with a transconjugant.