Help! Ovotransferrin impacts bacteria growth

[allison77]

Hi,

I wanted to test the antimicrobial effects of ovotransferrin on bacteria. Ovotransferrin works by binding to Fe3+ which most bacteria need. However, some bacteria need Fe2+ and some need Fe3+. I'm unsure where I should swab to get bacteria that will respond to the lack of Fe3+.

Additionally, since I don't have the proper equipment to extract ovotransferrin from egg white, I plan on doing a rescue experiment. I will use egg white, then use some iron source (would blood or rust work?) to see how the bacteria recovers. This is because there are other proteins in egg white that will kill the bacteria so I was thinking that supplying iron afterwards would promote the reproduction of the bacteria on the edge of the zones of inhibition. Then I could measure the amount of bacteria that "bounced back" by measuring the area? I'm still unsure of how the experiment should work however; I want to quantify (with numbers) a certain variable.

I was also thinking of measuring the catalase activity and how much gas is produced when mixed with H2O2 since lipid peroxidation depends on the ratio of Fe3:Fe2. But I'm unsure if this would actually work in real life? I would compare catalase activity before the egg white and then after the egg white.

Also wouldn't agar contain iron itself so wouldn't it saturate the ovotransferrin or is the iron amount insignificant?

I was also thinking of changing the agar to blood agar to on purposefully saturate the ovotransferrin to see how the bacteria would proliferate with extra iron.


I'm not exactly exactly sure how to format the procedure of the experiment so I am open to all suggestions, please reply!


Alison