Improved Production and Insulinotropic Properties of Exopolysaccharide by Phellinus igniarius in Submerged Cultures

Abstract

Phellinu igniarius (P. igniarius), a basidiomycete belonging to the family Polyporaceae, is a medicinal basidiomycetous fungus belonging to the Hymenochaetaceae and is an excellent remedy with anticancer and antioxidant qualities. The mushroom has been used as traditional medicines for the treatment of cardiovascular disease, tuberculosis, liver or heart diseases, bellyache, bloody gonorrhea, and diabetes. However, the limited production and market shortage have been attributed to the slow growth and the difficult collection of the fruiting body as well as the rare natural resources. The problem can be solved through the effective approach of submerged culture to produce a high bioactivity polysaccharide of P. igniarius. The project was proposed to investigate the effect of a surfactant on the production of polysaccharide in submerged culture of P. igniarius and their insulinotropic properties. Eight different surfactants including PEG series (4000, 6000), Tween series (20, 40, 80, 85), and Span series (20, 80) all at a concentration of 0.5 g/L were supplemented in turn to the basal medium in shake flasks. Among the various surfactants tested, Tween 80 exhibited the greatest exopolysaccharide production of 128.43 mg/L, and PEG 6000 showed the maximum biomass of 6.76 mg/mL. To find the optimal Tween 80 concentration for biomass and exopolysaccharide production, different Tween 80 levels (0, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2 g/L) were used in the medium. The maximal exopolysaccharide production of 132.76 mg/mL was achieved with the addition of 0.6 g/L of Tween 80 to the medium. The experimental results exhibited that the maximum of mycelia production in a stirred tank bioreactor was 3.01 mg/mL at Tween 80 0.2 g/L. In this study, their compounds, molecular weight, and protein content from fermentation product extracts were also tested. The average molecular weights of exopolysaccharide and intracellular polysaccharide were 1.715 × 106 Da and 4.87 × 105 Da, respectively. The protein contents of exopolysaccharide and intracellular polysaccharide were about 3.68% and 3.02%. The maximum RINm5F cell proliferations of exopolysaccharide and intracellular polysaccharide at 2 mg/mL were 142.3% and 120.07%, respectively. Cell proliferations of exopolysaccharide and intracellular polysaccharide increased with their concentrations. The maximum insulin secretion of exopolysaccharide at 2 mg/mL on RINm5F cell insulin was 0.615 μg/L.