With strong theoretical and practical significance, the principles and applications of enzymology are a key and difficult content in the course of biochemistry. In order to improve students’ comprehensive understanding and application ability of enzyme knowledge, a comprehensive experimental project “immobilization and isoenzyme identification of peroxidases” is designed. It integrates the main content of enzymes and focuses on five sections, namely, enzyme preparation, activity determination, characterization of enzymatic properties (Michaelis constant and stability), immobilization, and isoenzyme identification. Students’ ability to analyze and solve problems are improved by deep participation and coordinated implementation, and the experiment achieves a good teaching effect.