Abstract: To explore the potential for multiple uses of tobacco waste and the effects of extraction methods on the structure and function of polysaccharides, polysaccharides were extracted from tobacco waste by direct extraction and enzyme-assisted extraction. The process parameters of the enzyme-assisted extraction were optimized using response surface methodology, and the structure and antioxidant activity of the extracted polysaccharides were analyzed. The results showed that: 1) Under the conditions of a solid-liquid ratio of 1:32, cellulase addition rate of 0.455 g (enzyme activity of 5 000 U) per gram of tobacco waste, enzymatic hydrolysis temperature of 62 ℃, and enzymatic hydrolysis time of 2.0 h, the yield of polysaccharides was the highest, reaching 23.56%. 2) The polysaccharides obtained by the direct extraction and the cellulase-assisted extraction were predominantly acidic polysaccharides containing β-configuration pyran rings, with similar monosaccharide compositions. The polysaccharides obtained by the cellulase-assisted extraction showed a smooth, granular morphology with lower degrees of polymerization and cross-linking. 3) The polysaccharides from the both extraction methods showed some ability to scavenge DPPH and ABTS free radicals, indicating their antioxidant activity. This research provides theoretical support for the extraction, structural characterization, and bioactivity analysis for polysaccharides derived from tobacco waste, thus opening new possibilities for high-value utilization of tobacco waste.