Niuganba (NGB) is a traditional fermented beef product. Protease activity typically significantly affects the quality of NGB. Some natural food extracts may markedly influence NGB's protease activity and performance. This study aims to investigate the effect of Zanthoxylum bungeanum extract (ZBE) on the quality and cathepsin L activity of NGB. Following ZBE treatment, the myofibril fragmentation index (MFI), the content of TCA-soluble peptides, surface hydrophobicity, disulfide bond content, and cathepsin L activity of NGB significantly decrease. The content of free thiol groups and β-sheet significantly increases. Scanning electron microscopy (SEM) reveals that the arrangement of muscle fibers in the cross-section of NGB is more compact after ZBE treatment. The research results indicate that ZBE effectively inhibits cathepsin L activity, alleviates the degradation of myofibrillar proteins, improves the physicochemical characteristics of NGB, and enhances its structural stability.