<p>Subtilisin remains a key enzyme in enzymology due to its adaptability and broad applications. This review synthesizes recent progress from 2020 to 2025, integrating bibliometric data, structural insights, and modern applications. Our bibliometric analysis shows a rising interest in subtilisin research globally with Saudi Arabia and South Korea ranking prominently in publication productivity per million people (PPMP), underscoring their emerging leadership in this domain with a key shift toward sustainable production methods. This review highlights how using agricultural waste can reduce production costs by up to 60%, while promoting a circular economy. It also discusses how recent advancements in structural and computational biology provide a deeper understanding of the enzyme’s activity and stability, paving the way for targeted engineering. Subtilisin’s expanding uses in textile processing, waste management, and therapeutic development, including wound healing and thrombolysis have been explored. Looking ahead, opportunities lie in harnessing synthetic biology for tailor-made variants, employing AI-driven enzyme design for accelerated engineering, and translating subtilisin’s therapeutic potential into clinical applications. Despite this progress, challenges remain in optimizing its performance under harsh industrial conditions. This review aims to bridge foundational research with industrial innovation, offering a forward-looking perspective on subtilisin’s future biotechnological potential.</p> Graphical Abstract <p></p>