This review summarizes the latest research on nanomaterials for co-immobilization of multiple enzymes over the past five years. Enzymes are essential biocatalysts with high specificity and efficiency, but their industrial application is limited by instability under harsh conditions. Enzyme immobilization enhances stability, reusability, and operability, and is achieved through various methods such as physical adsorption, encapsulation, covalent bonding, and cross-linking. Multi-enzyme cascade reactions offer advantages over single-enzyme reactions by eliminating the need for separation and purification steps, improving substrate conversion efficiency, and minimizing side reactions. Nanomaterials, with their tunable morphology, high surface area, and abundant active sites, are ideal for enzyme co-immobilization. They improve enzyme stability, activity, and catalytic efficiency, and provide a stable environment for enzyme reactions. The review discusses various nanocarriers, including metal-based nanoparticles, magnetic nanoparticles, metal-organic frameworks, mesoporous silica nanoparticles, and carbon-based nanomaterials, and their advantages and challenges in enzyme co-immobilization. The review also highlights the potential of these nanomaterials in applications such as biosensing, biocatalysis, medical diagnostics, and therapeutics. The study emphasizes the importance of optimizing immobilization conditions, selecting appropriate carriers, and designing effective immobilization strategies to enhance enzyme activity and stability. The review concludes that nanomaterials offer great potential for efficient and selective chemical transformations through multi-enzyme co-immobilization.This review summarizes the latest research on nanomaterials for co-immobilization of multiple enzymes over the past five years. Enzymes are essential biocatalysts with high specificity and efficiency, but their industrial application is limited by instability under harsh conditions. Enzyme immobilization enhances stability, reusability, and operability, and is achieved through various methods such as physical adsorption, encapsulation, covalent bonding, and cross-linking. Multi-enzyme cascade reactions offer advantages over single-enzyme reactions by eliminating the need for separation and purification steps, improving substrate conversion efficiency, and minimizing side reactions. Nanomaterials, with their tunable morphology, high surface area, and abundant active sites, are ideal for enzyme co-immobilization. They improve enzyme stability, activity, and catalytic efficiency, and provide a stable environment for enzyme reactions. The review discusses various nanocarriers, including metal-based nanoparticles, magnetic nanoparticles, metal-organic frameworks, mesoporous silica nanoparticles, and carbon-based nanomaterials, and their advantages and challenges in enzyme co-immobilization. The review also highlights the potential of these nanomaterials in applications such as biosensing, biocatalysis, medical diagnostics, and therapeutics. The study emphasizes the importance of optimizing immobilization conditions, selecting appropriate carriers, and designing effective immobilization strategies to enhance enzyme activity and stability. The review concludes that nanomaterials offer great potential for efficient and selective chemical transformations through multi-enzyme co-immobilization.