This study aimed to investigate the physicochemical properties, health-promoting benefits and rheological properties of an EPS produced by a novel probiotic Pediococcus pentosaceus M41 isolated from a marine source. P. pentosaceus M41 was able to produce an EPS with average molecular weight of 682.07 kDa. EPS-M41 consisted of arabinose, mannose, glucose and galactose with a molar ratio of 1.2:1.8:15.1:1.0. EPS-M41 structure could be proposed as →3)α-D-Glc(1→2)β-D-Man(1→2)α-D-Glc(1→6)α-D-Glc(1→4)α-D-Glc(1→4)α-D-Gal(1→ with arabinose linked at the terminals. At concentration of 10 mg.ml −1, the antioxidant capacity was 76.5% and 48.9% for DPPH and ABTS, respectively. EPS-M41 inhibited 86.8% and 90.8% of the α-amylase and α-glucosidase activities, respectively, at 100 μg.ml −1. A 77.5% and 46.4% of antitumor inhibition occurred by EPS-M41 against Caco-2 and MCF-7 cells, respectively. The apparent viscosity (ƞ) of all EPS-M41 solutions decreased with shear rate increases. Salt type and pH value had an impact on the rheological properties of EPS-M41.