Three-Phase Partitioning for the Extraction and Purification of Polysaccharides from the Immunomodulatory Medicinal Mushroom Inonotus obliquus

Abstract

Polysaccharides from the immunomodulatory medicinal mushroom Inonotus obliquus (IOPS) were extracted and purified using three-phase partitioning (TPP), which is an efficient, fast, safe, and green purification technique. An optimal extraction procedure that gave a good 2.2% isolated yield was identified, using the following protocol: a solid-liquid ratio of 1 g to 12 mL; mass fraction of (NH4)2SO4 20% (w/v); 11 mL t-butanol; pH 8.0; temperature 30 °C; and extraction time 30 min. The purified IOPS was shown to be a proteoglycan of 40 kDa molecular weight comprising of d-galactose, d-glucose, d-xylose, and d-mannose in a molar ratio of 2.0:3.5:1.0:1.5. The purified IOPS displayed strong free-radical scavenging abilities, antioxidant activities, and immunological activity in vitro. IOPS’ Trolox antioxidant equivalent capacity and ferric-reducing ability of plasma were 251.2 μmol Trolox/g sample and 1040.5 μmol Fe2+/g sample, respectively, with the activity of its immunomodulatory behavior shown to be gradient dependent.

Global optimization for simultaneous extraction of oil and polysaccharides from Schizochytrium limacinum by enzyme‐assisted three‐phase partitioning

“…When the mass fraction of ammonium sulfate was 30%, the recovery rate of polysaccharides and oil reached the maximum. However, as the mass fraction of ammonium sulfate continued to increase, the hydrogen bond network formed between the polysaccharides, and the water molecule was weakened; moreover, and the network was destroyed, thereby reducing the polysaccharides extraction rate (Liu et al., 2019). The continuous increase in the mass fraction of ammonium sulfate increases the ionic strength of the mixture, thereby leading to the decomposition of triglyceride into carboxylic acid and glycerol and reducing the oil yield (Dutta, Sarkar, & Mukherjee, 2015).…”

“…I G U R E 1 Effect of enzyme concentration (a), and enzymolysis time (b) on the yield of Oil and polysaccharides Effect of different mass fraction of ammonium sulfate (a), ratio of slurry to t-butanol (b), time (c) and temperature (d) on the yield of oil and polysaccharides extraction rate(Liu et al, 2019). The continuous increase in the mass fraction of ammonium sulfate increases the ionic strength of the mixture, thereby leading to the decomposition of triglyceride into carboxylic acid and glycerol and reducing the oil yield(Dutta, Sarkar, & Mukherjee, 2015).…”

Recent Developments in Inonotus obliquus (Chaga mushroom) Polysaccharides: Isolation, Structural Characteristics, Biological Activities and Application

“…As most polysaccharides have good water solubility, the classic method uses hot water reflux to extract polysaccharides, Gao determined the optimal process for traditional water extraction by orthogonal experiments, 80 °C for temperature, 1.5 h for time, 1:40 ( w / v ) for material-to-liquid ratio, the yield for IOPS is 2.53% [ 65 ]. Despite that this method is simple and easy to implement, the efficiency was very low, the yield of IOPS varies from 2.12% to 2.53% [ 15 , 66 ].…”

Bioactive compounds and bioactive properties of chaga ( Inonotus obliquus) mushroom: a review

“…For example, the purified chaga polysaccharide reported by Liu et al (2019), is a proteoglycan with a MW of 40 kDa and contains 57.17% carbohydrate and 32.53% protein. The carbohydrate part is comprised of D-galactose, D-glucose, D-xylose, and D-mannose in a 2.0:3.5:1.0:1.5 mole ratio.…”