Abstract
Everyone is in constant search for the next anti-aging solution, from skincare to supplements, but could social behavior itself influence the rate of aging? Studies in humans and animals suggest that positive social interactions correlate with longevity, while negative social environments are linked to accelerated aging. In Drosophila melanogaster, aggression is a well-studied behavior regulated by genetic and environmental factors. One gene of interest, Cyp6a20, has been identified as a modulator of aggression. This gene encodes a cytochrome P450 enzyme, but its functional role remains unexplored. This study aimed to express and purify Cyp6a20 protein, laying the groundwork for future biochemical and structural analyses. Our findings show that Cyp6a20 knockout (KO) flies (loss of Cyp6a20 function) displayed increased aggression along with sex-specific changes in pheromones. These behavioral and pheromone changes were associated with reduced lifespan, suggesting a role for Cyp6a20 in linking social behavior and aging. Iterative optimizations to the expression and purification method led to the development of an effective Cyp6a20 purification protocol. Construct enhancements, including the addition of a linker sequence, significantly improved purification results. Additionally increasing the concentration of 5-aminolevulinic acid (5-ALA) during expression ensured heme incorporation and resulted in better Cyp6a20 expression. These optimizations provide a reliable expression and purification method, laying the foundation for future studies of the enzyme’s biochemical activity and its role in pheromone signaling and aging.