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Team:Acton-BoxboroughRHS/Project
From 2014hs.igem.org
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ABRHS iGEM Site 
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Kopi Luwak coffee is the rarest and the most expensive type of coffee in the world. This beverage is made from the feces of the Asian Palm Civet (Paradoxurus hermaphroditus) or Luwak, a cat-like omnivore that fills the niche of a racoon in Asia. The Asian Palm Civet eats raw coffee berries. As the berries are digested, enzymes in the animal's digestive tract break down components of the coffee bean that are responsible for its bitter taste. However, the beans themselves are not digested. The civet only digests the fleshy outer layer, so when it defecates, it leaves clumps of coffee beans that have been processed by its enzymes. The beans are then cleaned, roasted, and brewed to make the Kopi Luwak coffee.
Due to the complexity of this process, Kopi Luwak is a very expensive item at $600 per pound. The outrageous price has made Kopi Luwak a novelty for the rich. Its allegedly excellent flavor is sadly something that most people are unable to afford. Another drawback of Kopi Luwak is that demand has driven businesses to animal cruelty in order to keep up production. A small civet farming industry has tens of thousands of civets living in battery cages being force fed coffee berries. Civets, being omnivores, are no more capable of surviving on coffee than humans. As a result their population is diminishing.
We propose to make this process more humane, efficient, and sanitary by using bacteria instead of civets to process coffee berries into Kopi Luwak beans. We will accomplish this by inserting genes that code for proteins found in the civet's digestive tract into a hardy bacteria that can withstand the pH levels required for the proteins to operate. We intend to add genes for salivary amylase, pepsin, pancreatic amylase, trypsin, and chymotrypsin, sequenced from palm civets or closely-related species. Once the bacteria have been transformed, we will attempt to simulate the digestive process of the civet on coffee berries and analyze our results.
Although this project seems unconventional, putting animal enzymes in bacteria has been done before to great effect. Take rennet cheese for an example. In order to obtain the enzymes required to produce this cheese, a calf must be slaughtered; the material (rennet) is taken from the dead animal's digestive tract. Now, due to growing demand for this cheese, bacteria containing the rennet enzymes are used instead. Today, the sale of these cheeses is a popular and profitable industry. By putting animal enzymes in bacteria, we create a digestive platform that is capable of processing more than just coffee berries.
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