Development and implementation of a project-based laboratory module: heterologous expression and fermentation of ergothioneine in E. coli for undergraduate biochemistry and molecular biology education

There is a growing need in biochemistry education to move beyond traditional, recipe-based experiments and toward integrated, research-based experiences that mimic real-world scientific inquiry. We designed a multi-week, project-based learning (PBL) module where students undertake the complete process of producing ergothioneine (ERG), a valuable antioxidant, using recombinant DNA technology in Escherichia coli . The project encompasses molecular cloning (optional), protein expression analysis, fermentation in a bioreactor, and product quantification. The module was successfully implemented in an upper-level biochemistry laboratory course. Assessment via pre- and post-lab quizzes, lab reports, and a comprehensive student self-evaluation survey showed significant gains in technical skills, conceptual understanding, and research confidence. This laboratory module provides an effective framework for integrating modern biotechnological techniques into the undergraduate curriculum. It enhances student learning by contextualizing theoretical knowledge within a continuous, authentic research project, thereby fostering critical thinking, problem-solving skills, and enthusiasm for biochemical research.