Purification of polysaccharides from Phellinus linteus by using an aqueous two-phase system and evaluation of the physicochemical and antioxidant properties of polysaccharides in vitro.

In this study, Phellinus linteus polysaccharides (PLPS) and proteins were simultaneously separated from P. linteus mycelia by using an aqueous two-phase system (ATPS) based on choline chloride ([Chol]Cl)/K2HPO4, and the physicochemical and antioxidant properties of PLPS after ATPS extraction were evaluated. Results demonstrated that the maximal extraction efficiencies of 68.53% ± 0.29% PLPS and 82.37% ± 0.41% proteins were obtained when the cholinium-based ATPS contained 68.9% K2HPO4, 20% [Chol]Cl, 10.0 mg mL-1 crude water extract (1.0 mL), and distilled water (4.0 mL) at shaking time and temperature of 30 min and 21.2 °C, respectively. Compared with C-PLPS obtained using traditional ethanol precipitation and isolation protocols, PLPS had higher carbohydrate content (63.58% ± 1.12%), lower molecular weight (15.2 kDa, 80%), different monosaccharide compositions, and showed similar preliminary structural characterizations. Moreover, PLPS exhibited more evident scavenging effects on free radicals and in vitro antioxidant activities than C-PLPS. Therefore, the method of [Chol]Cl/K2HPO4 ATPS can be developed as an effective strategy for the separation/purification of highly bioactive polysaccharides.