Evaluating Probiotic Protein Expression as a Function of Oxidation Over Time

Anjul Bansal, Grady Blacken, HU Chemistry Department, Bellevue College, 3000 Landerholm Cir SE, Bellevue, WA 98007

Probiotics are found in numerous fermented foods such as yogurt, sauerkraut, and kombucha. Research shows that eating live probiotics are beneficial for the gut because it helps supplement the plethora of native bacteria. Now, these microorganisms are gaining popularity throughout the world—people are ingesting them in the form of food, drink, and even pills. Most probiotics are anaerobic. In fact, excess oxygen can damage organelles, create ionic imbalance and eventually even kill the cells. However, in the process of manufacturing these fermented foods, the probiotics in them often get exposed to oxygen, such as due to leaks in packaging. Even the FDA does not have a rule about standard manufacturing processes regarding anaerobic conditions and yogurt. This could mean that the probiotics people eat are highly compromised: damaged or dead.

 We hypothesize that probiotics exposed to aerobic environments for extended periods of time will express more DNA repair enzymes such as DNA pol. 1 and 2, p53 or photolyase. This is because oxygen will cause probiotic cells to change function, and therefore they will start to express enzymes, such as these DNA repair enzymes, to protect them from oxidative damage. In order to test this, probiotics will be left out to oxidize for various amounts of time—0 minutes, 1 hour, 2 hours, and 24 hours. After oxidizing, the cells will be cultured for 16 hours. Cells will then be pelleted by centrifugation at 300 x g and washed and lysed. Then, cell proteins will be tryptically digested and analyzed by LCMS (Liquid Chromatography Mass Spectrometry) for identification.

In the future, these proteins can provide insights to how these oxidized probiotics impact the gut. Are they really the miracle microorganisms that benefit the gut, or could their compromised condition end up being harmful?

Additional Abstract Information

Presenter: Anjul Bansal

Institution: University of Washington, Seattle

Type: Poster

Subject: Biochemistry

Status: Approved

Time and Location

Session: Poster 2
Date/Time: Mon 3:00pm-4:00pm
Session Number: 2512