Network Analysis of Drug- and Dye-Modifying Azoreductase Enzymes Found in the Gut Microbiome
Network Analysis of Drug- and Dye-Modifying Azoreductase Enzymes Found in the Gut Microbiome
Brisa Mendoza ’24, Biochemistry major
Emma Mortara ’24, Biochemistry major
Faculty Mentor: Dr. Tyler Stack, Chemistry and Biochemistry
Poster Presentation: Wednesday, April 24, 1:30 – 3 p.m.
Individual patients have diverse and dynamic gut microbiomes that can metabolize drugs, affecting therapeutic efficacy and safety. In the human gut microbiome, xenobiotic metabolism is exemplified by the reduction of the azo bond (R1-N=N-R2) of certain drugs and food additives by enzymes characterized as azoreductases. Using a sequence similarity network of an azoreductase protein family, 24 representative sequences found in abundance in the human gut microbiomes were prioritized and their activity with a group of azo compounds were analyzed. A representative azoreductase (UniProt ID: A0A1M6LNR3) was found to reduce the azo drugs sulfasalazine and 3-phenylazo-2,6-diaminopyridine monohydrochloride, the azo-dye methyl red and a non-azo dye indigo carmine. We used enzyme kinetics to characterize this enzyme for its NADH-dependent reduction of these drugs and food additives. Nuclear magnetic resonance and mass spectrometry were then used to confirm reaction completion and identify product compounds. Overall, this strategy aims to minimize the number of unique azoreductases needed to characterize one protein family in the diverse set of potential drug- and dye-modifying activities in the human gut microbiome. Characterizing the activity of azoreductases in the gut microbiome can assist in developing more effective therapeutics that reduce side effects and move towards personalized medicine.
