The structure and hypoglycemic activity of tea polysaccharides has been extensively studied, while there are few reports on the characterization and hypoglycemic activity of dark tea polysaccharides. The crude dark tea polysaccharide (CDTPS) was optimally extracted from Fuzhuan dark tea. Six polysaccharide fractions (namely DTPS-1, DTPS-2, DTPS-3, DTPS-4, DTPS-5, and DTPS-6) were isolated from CDTPS, and their physicochemical, structural, and biological properties were compared and analyzed. The results revealed that the compositions, structural characteristics, and biological properties of the six DTPSs were different. Therein, DTPS-4 and DTPS-6 had looser morphology, faster solubility, and a more stable structure. Additionally, DTPS-4 had the optimum in vitro antioxidant capabilities, and DTPS-6 had the strongest in vitro hypoglycemic capabilities. In addition, a correlation analysis revealed that the molecular weight and uronic acid content were significantly related to their antioxidant and hypoglycemic activities. Our results indicated that DTPS-4 and DTPS-6 could be further developed into functional foods or additives, respectively.