A novel heteropolysaccharide (GPP-TL) was isolated from tetraploid Gynostemma pentaphyllum (Makino) leaf by hot water extraction and anion-exchange and gel permeation chromatography approaches. GPP-TL had a molecular weight of 9.3 × 10(3) Da and was primarily composed of glucose, galactose, and arabinose, with a molar ratio of 43:5:1, respectively. The chemical structure of GPP-TL was characterized using chemical and instrumental analyses. The results indicated the presence of (1→4)-α-d-glucopyranosyl, (1→4)-β-d-galactopyranosyl, (1→4,6)-linked-α-d-glucopyranosyl, and terminal 1→)-α-d-glucopyranosyl moieties in a molar ratio of 5.7:1:1.5:1, respectively. The results indicated that GPP-TL had glucose and galactose residues in the main chain with (1→6)-linked branches at glucose residues. In addition, GPP-TL exhibited scavenging capacities against hydroxyl, peroxyl, and DPPH radicals in vitro and had a stronger bile acid-binding ability than psyllium on a same-weight basis.