 Nanomaterials are used to enhance enzyme activity through substrate channeling while improving enzyme stability and reusability in multi-enzyme cascade reactions, which have become a focus of intensive research due to significant debates surrounding aspects such as enzyme orientation, loading, retention, and immobilization techniques. Researchers have designed functional in vitro multi-enzyme systems inspired by natural processes within living organisms, achieving substantial progress in designing multi-step reactions that harness the synthetic capabilities of various enzymes in applications such as biomarker detection and biofuel cells. This review provides an overview of recent developments in concurrent and sequential approaches involving two or more enzymes in sequence on nanostructured electrodes, addressing challenges encountered and innovative solutions devised in this field. This article was authored by Shalini Devi Kalyana Sundram, M.D. Motahar Hossain, Mohamed Resky, and others. We are article.tv. Links in the description below.