1. Genesis of Mankind and Food The Ultimate Bacteria that Exterminate Food Poisoning by Aflatoxins *The background theme of this PPT is unrelated to my project.

2. 1 in every 6 Americans suffer from food poisoning annually. Fungi that grow on moldy breads and other moldy foods produce mycotoxins, which are some of the most deadly biotoxins in the world. Fun fact: mycotoxins can cause cancer, liver and kidney diseases, lots of pain, and even death.

3. This project is focused on a type of mycotoxin called aflatoxin, the most dangerous mycotoxin that serves as a carcinogen. The goal is to engineer a species of bacteria that is able to destroy the aflatoxin protein. The bacteria will live symbiotically in our guts, therefore they are able to reproduce while containing the gene to destroy the toxin proteins.

4. An array of the harmless strain of Escherichia coli bacteria from our guts to serve as the main power for destroying the aflatoxin proteins. Promoter linking to the immunogen MAI-16885, used to sense and activating an immune response Acidic protease gene sequence from the bacteria species Saccharomycopsis fibuligera, to dissolve the aflatoxin proteins Buffer gate

5. The bacteria E. coli are chosen for their symbiotic abilities and their gram-negative quality, which allows the bacteria to have a membranous (gel-like) plasmic membrane. This is an image of gram- negative stain of E. coli. Because the bacteria are gram-negative, their gram stain is violet-colored.

6. The Design - Continued The aflatoxins release a substance called AFB1- guanine adduct inside of the intestine. In order for the bacterial cells to detect this substance, we need a promoter, which is sensitive to the AFB1-guanine adduct, that is linked to the immunogen MAI-16885.

7. The Design - Continued In order for the bacterial cell to destroy a toxin protein, an acidic protease* gene sequence from the yeast species In order for the bacterial cell to destroy a toxin protein, an acidic protease* gene sequence from the yeast species Saccharomycopsis fibuligera is required to be inserted into the E. coli bacteria’s plasmids, so they can obtain the ability to produce this acidic protease. The yeast Saccharomycopsis fibuligera The acidic protease *The acidic protease would then need to be further engineered to become specifically fit by aflatoxin’s tail end structure of –OCH3.

8. The Design - Continued A Buffer gate is required. When the AFB1-guanine adduct is present, the process begins. When the AFB1-guanine adduct no longer exists in the guts, the process stops. For a more specific Buffer gate table, please visit my Wikispaces page at: http://bli-biotech-research.wikispaces.com/Jim

9. The promoter detects the AFB1-guanine adduct, and activates the MAI-16885 in order to trigger an immune response from the bacterial cell. I TRIGGER THOU! This light doe.

10. PoE - Continued Because E. coli bacteria are gram-negative, they are able to make vesicles from their plasmic membranes and send them to other places. The bacterial cell then produces an acidic protease from its ribosome, and a vesicle is sent to capture the acidic protease. The vesicle reaches the surface of the cell, and forms another vesicle carrying the same protease to the outside of the cell.

11. PoE - Continued The promoter sends information to the cell about the direction of the aflatoxin protein. The vesicle travels to a nearby aflatoxin protein. The vesicle captures the aflatoxin protein, in which the acidic protease begins a denaturation of the aflatoxin protein. Diagram on the next slide. Please enjoy this luxury.

12. Acidic Protease Aflatoxin Protein

13. Promoter detecting AFB1- guanine adduct Bacteri a MAI-16885Immunogen AFB1-guanine adduct Ribosomes Acidic Protease Vesicle Aflatoxin

14. Medications for aflatoxin poisoning are rare, and they are usually used in emergency situations. I want a patient to be cured BEFORE the situation becomes an emergency. Most tests to detect the presence of aflatoxin take a long time before results come out, usually around twenty-four hours.

15. Thank so much for listening! Now go enjoy your life. And I pray that my idea will come to life one day.