The gut microbiome is composed of trillions of microorganisms that play a large role in host health and disease. There is evidence that the composition of the gut microbiome differs depending on sex. In the gut microbiome, there are specific microbes that contain β-Glucuronidase enzymes. These enzymes deconjugate host-derived molecules, such as sex steroids and bile acids, and deconjugate external compounds, such as xenobiotic conjugates. The goal of this research is to determine whether sex or Hpg axis affects glucuronidase activity in the gut microbiome. Fecal samples were collected from four groups of mice: female and male wild type mice that undergo puberty, and female and male Gnrh1hpg mutant mice that do not. We also used bioinformatics analysis of genomes in our gut bacterial culture collection to identify those with potential β-Glucuronidase activity. Glucuronidase and Bradford assays will be performed on fecal samples and bacterial cultures, and statistical analyses (ANOVAs) will be used to detect differences in activity in murine fecal samples and bacterial cultures. It is evident that there are sex-related differences in the composition of the gut microbiome, however the reasons for these differences, and the impact of these differences on the host is not yet understood. One possible mechanism involves sex steroids that enter the gut and are deconjugated by specific bacteria, changing the overall composition of the gut microbiome. Testing for β-Glucuronidase activity in these samples will allow us to compare activity levels related to sex and Hpg axis, giving us a better understanding on the mechanism of how sex and sex steroids influence the gut microbiome. In future research, this could allow for the development of sex-specific treatment options, depending on β-Glucuronidase levels of a patient.