"
Team:UIUC Illinois/Project/PuppyProbiotic
From 2014.igem.org
| Line 23: | Line 23: | ||
<div class="rightparagraph"> | <div class="rightparagraph"> | ||
<p><b>What just happened?</b> | <p><b>What just happened?</b> | ||
| - | <br>Canines and many small animals are incapable of metabolizing theobromine and related stimulants as fast as humans can. | + | <br>Canines and many small animals are incapable of metabolizing theobromine and related stimulants as fast as humans can. Chocolate contains theobromine, an alkaloid that has stimulant properties. An excess of the stimulant builds up and begins to overstimulate the animal, thus leading to many malicious side effects. Ranging from slight nausea to death, consumption of these compounds are harmful for the animal. Chocolate can affect the animal regardless of the amount administered. Even the smallest amounts can produce side affects like diarrhea. In fact, theobromine has been employed to ward off pests of the same genus as canines: "Results indicate that mixtures of theobromine and caffeine have the potential to be developed into a selective, effective, and socially acceptable toxicant for the control of pest coyotes."[1] |
| + | <img src="https://static.igem.org/mediawiki/2014/7/7a/Theobromine-molecule.gif" alt="Smiley face" width="500" height="650"> | ||
</div> | </div> | ||
<div style="clear: both;"></div> | <div style="clear: both;"></div> | ||
| Line 47: | Line 48: | ||
<br>UT Austin developed this part by rearranging the operon of CBB5 and developed this part: | <br>UT Austin developed this part by rearranging the operon of CBB5 and developed this part: | ||
<img src="https://static.igem.org/mediawiki/2012/2/25/Austin_Texas_Decaffeination_design.png" alt="Smiley face" width="500" height="100"> | <img src="https://static.igem.org/mediawiki/2012/2/25/Austin_Texas_Decaffeination_design.png" alt="Smiley face" width="500" height="100"> | ||
| + | <br>By using this construct, we were eager to repurpose it to degrade theobromine. | ||
</p> | </p> | ||
Revision as of 02:22, 17 October 2014
Puppy Probiotics.
An Introduction
Imagine... Imagine drinking a cup of coffee. You stolidly sip your espresso and soon enough you feel a warm sensation cajole your body into getting that assignment done. However, you notice you're fidgeting uncontrollably, the room starts spinning and you begin to vomit. Ambulances, the noises are familiar, you've just seized. You're a dog.
What just happened?
Canines and many small animals are incapable of metabolizing theobromine and related stimulants as fast as humans can. Chocolate contains theobromine, an alkaloid that has stimulant properties. An excess of the stimulant builds up and begins to overstimulate the animal, thus leading to many malicious side effects. Ranging from slight nausea to death, consumption of these compounds are harmful for the animal. Chocolate can affect the animal regardless of the amount administered. Even the smallest amounts can produce side affects like diarrhea. In fact, theobromine has been employed to ward off pests of the same genus as canines: "Results indicate that mixtures of theobromine and caffeine have the potential to be developed into a selective, effective, and socially acceptable toxicant for the control of pest coyotes."[1]
What can we do?
Problem solving
In effort to solve this problem, our team looked at the biological cascade of caffeine and theobromine. Since our goal was to degrade caffeine into something less harmful, we looked to digestion. We uncovered that theobromine can in fact be degraded into subtituents that are less harmful, namely a cascade that converts it to uric acid, a molecule that the dogs could excrete as waste
Nature's Answer For all techniques listed, our lab required usage of gloves, goggles, lab coats, closed-toe shoes and long pants to minimize the amount of contact between us and the materials that are being used. Also, general aseptic protocols were adhered to when handling any lab techniques to prevent contamination. Our lab required us to go through a live training with a lab representative to learn proper handling and care of the autoclave.
Nature's answer We discovered that the organism Pseudomonas putida CBB5 endogenously metabolized caffeine and subsequent methylxanthines. In fact, the organism degraded the compounds using the proteins NdmA (N-Demethylase A) and NdmB, both of which are Rieske nonheme iron monooxygenases, NdmC, another demethylase and NdmD, a reductase. By using this pathway, the organism is able to use caffeine as a carbon and nitrogen source.[2] Too good to be true? No. In fact, a previous iGEM team, UT Austin 2012 Caffeinated Coli, harnessed the power of this novel pathway in their own project! We were ecstatic to learn that the pathway was available in biobrick form and ready for us to manipulate.
UT Austin developed this part by rearranging the operon of CBB5 and developed this part:
By using this construct, we were eager to repurpose it to degrade theobromine.
illinoisigem@gmail.com 
