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Caffeine

Caffeine is arguably the most popular stimulant and psychoactive substance consumed in recent times. This is evident from the huge demand for coffee and tea beverages and increasingly energy drinks, all of which are common dietary sources of caffeine (Johnson, 2011). Currently, methods of isolating caffeine for use as an additive include the decaffeination of commercially harvested plants, such as green tea leaves and Coffea arabica beans. This is often an expensive process involving large quantities of organic solvents or water, or specialized techniques such as supercritical fluid extraction with carbon dioxide (Vuong & Roach, 2013).

The ecological and financial burden of current caffeine production methods makes a synthetic biology approach desirable. While there has been significant research done on the degradation of caffeine using microbes (Lakshmi & Das, 2011; Asano et al., 1993), synthesizing caffeine in a biological chassis is less common in the literature. Our goal was to implement the caffeine biosynthetic pathway in E. coli to enable the cost-effective production of caffeine in a bioreactor. This is largely building on the work of the 2012 TU Munich team, which successfully got caffeine precursors expressed in yeast.

Tying in with our overall goal of immunizing bioreactors using the CRISPR system, the caffeine biosynthetic pathway serves as part of a proof-of-concept showing that the CRISPR system could be used to alter population dynamics in bioreactors.

Sources:
Asano, Y., Komeda, T., & Yamada, H. (1993). Microbial production of theobromine from caffeine. Bioscience, Biotechnology and Biochemistry, 57(8), 1286-1289. Retrieved from https://www.jstage.jst.go.jp/article/bbb1992/57/8/57_8_1286/_article
Johnson, B. A. (2011). Addiction medicine. (Vol. 1, pp. 551-583). New York, NY: Springer Science Business Media, LLC. Retrieved from http://link.springer.com/chapter/10.1007/978-1-4419-0338-9_26
Lakshmi, V., & Das, N. (2011). Biodegradation of caffeine by trichosporon asahii isolated from caffeine contaminated soil. International Journal of Engineering Science and Technology, 3(11), 7988. Retrieved from http://connection.ebscohost.com/c/articles/79461823/biodegradation-caffeine-by-trichosporon-asahii-isolated-from-caffeine-contaminated-soil
Vuong, Q. V., & Roach, P. D. (2013). Caffeine in green tea: Its removal and isolation. Separation & Purification Reviews, 43(2), 155-174. Retrieved from http://www.tandfonline.com/doi/full/10.1080/15422119.2013.771127