Abstract Surface-Enhanced Raman Scattering (SERS) is an analytical test technique that uses metal or metal nanoparticles as a detection substrate. It can be used to characterize molecular vibration information with good reproducibility and stability. Nanozymes are nanomaterials with catalytic functions. In recent years, the research on the catalytic activity of nanozymes has developed rapidly, which has aroused extensive research interest in biology and medicine. Nanozymes can avoid the characteristics of easy inactivation of biological enzymes, exhibits high stability and good catalytic performance in water or buffer solution. It has broad application prospects in the field of catalysis and enzyme kinetics due to its catalytic activity and simple preparation method. However, the research on the combination of SERS technology and the simulation of the catalytic activity of biological enzymes is limited. Most of the studies use UV-visible absorption spectroscopy to analyze the catalytic performance of nanozymes. In this paper, Ag nanoparticles in the PANI matrix were prepared by one-step self-assembly redox polymerization. In the polymerization of aniline, a composite of AgNO3-(3-aminopropyl) triethoxysilane (APTES) acts as oxidants and structure inducers; then the reduction of AgNO3 is accompanied by the oxidation of aniline, resulting in the formation of Ag nanoparticles within a PANI matrix. It can be found that nanocomposite can function as not only peroxidase and glucose oxidase, but also tandem enzyme, which directly reflect glucose concentration by oxidizing TMB. Therefore, SERS technology and simulated enzyme catalysis research are combined in this paper, and SERS technology is used to detect H2O2, glucose and TMB more quickly and effectively.