This study extracted and purified a natural polysaccharide (TPS-5) that has a molecular weight of 48.289 kDa from Liupao tea, a typical dark tea with many benefits to human health. TPS-5 was characterized as a pectin-type acidic polysaccharide. It has a backbone composed of → 2,4)- α- L-Rhap-(1) → 4)- α- D-GalAp-(1) →, with a branch composed of → 5)- α- L-Ara-(1 → 5,3)- α- L-Ara-(1 → 3)- β- D-Gal-(1 → 3,6)- β- D-Galp-(1) →. The in vitro biological activity evaluation illustrated that TPS-5 has free radical scavenging, ferric-ion-reducing, digestive enzyme inhibitory, and bile-salt-binding abilities. These results suggest that TPS-5 from Liupao tea has potential applications in functional foods or medicinal products.