The physicochemical properties of the polysaccharides in Polygonatum kingianum, a Chinese medicinal herb used for both medicine and food, have not been fully studied. This study isolated three polysaccharides (PKP-1, PKP-2, and PKP-3) from the dry rhizomes of P. kingianum, with an average molecular weight of approximately 3137 Da, 5341 Da and 3755 Da, respectively. Structural analysis showed that all the three polysaccharides are fructans with β-D-Fruf-(2→, →6)-β-D-Fruf-(2→, →1)-β-D-Fruf-(2→, →1,6)-β-D-Fruf-(2→ and →6)-α-D-Glcp-(1→ glycosidic bond type. Notably, PKP-2 contains both acetyl groups and trace amounts of mannose residues. Scanning electron microscopy indicated that each polysaccharide possesses unique surface morphology. Thermal analysis showed that the three polysaccharides have good thermal stability. Rheological studies further revealed that all the three polysaccharides are typical shear thinning fluids. In vitro experiments showed that PKP-1 and PKP-2 significantly promote the secretion of NO and cytokines (TNF-α, IL-6) in macrophages by activating the NF-κB signaling pathway, thereby demonstrating potential immunomodulatory activity. These findings lay a theoretical foundation for the potential application of Polygonatum polysaccharides in the food industry.