Proteases play crucial roles in the life cycles of bacterial, fungal, and viral pathogens, making them prime targets for therapeutic intervention. Screening for inhibitors of these proteases is essential for developing novel treatments for infectious diseases. This chapter explores the use of Saccharomyces cerevisiae (baker’s yeast) as a model system in growth restoration screens to identify inhibitors of pathogenic proteases. Yeast offers a highly adaptable platform due to its ease of genetic manipulation, rapid growth, and cost-effectiveness, allowing the expression of heterologous proteases from bacterial, fungal, and viral sources. In growth restoration screens, S. cerevisiae cells are engineered to express toxic proteases from pathogens, leading to impaired growth. The introduction of protease inhibitors restores yeast cell growth, providing a straightforward and scalable readout for identifying potential drug candidates. This chapter outlines the methodology for cloning and expressing proteases in yeast, employing inducible promoters for controlled expression, and using small molecule libraries to screen for inhibitors. Key examples are provided where this approach has successfully identified protease inhibitors for bacterial (e.g., Staphylococcus aureus ClpP), fungal (e.g., Candida albicans aspartic proteases), and viral (e.g., HIV, SARS-CoV-2) targets. The chapter also discusses the challenges of protease expression in yeast, issues with false positives, and the need for secondary assays to validate inhibitor specificity. Despite these limitations, S. cerevisiae-based screens have proven effective for high-throughput drug discovery, with significant potential for targeting a wide range of proteases. In future, this approach will continue to play a critical role in developing therapeutic strategies against emerging and resistant pathogens. This versatile screening platform underscores the importance of yeast models in modern drug discovery and their contribution to addressing global health challenges.