Implications of Ultrasonication-assisted Extraction with Response Surface
Methodology on Phytochemical Compositions and Antioxidant Activity of
Polysaccharide Extract from Phellinus rimosus (Berk.) Pilát Cultivated
Mycelia in Northeastern Thailand
Authors: Maingam Chonlada , Kanchanarach Watchara , Chutiman Nipaporn , Wanthong Anuwat , Srivilai Prayook , Loutchanwoot Panida 
Published: February 2024

Background:
Polysaccharides from the medicinal mushroom Phellinus rimosus (Berk.)
Pilát (PR) are the major functional bioactive ingredients. However, there has been a marked natural
decrease in the number of PR fruit bodies, leading to their increased cost. Moreover, the natural
growth and development of mature PR fruit bodies takes several decades.

Objective:
The objective of this study was to produce a polysaccharide extract from cultured PR
mycelia (PEPRM) by using ultrasonic-assisted extraction with response surface methodology (RSM),
and determine its physicochemical composition and antioxidant potential.

Methods:
Polysaccharide and monosaccharide composition analyses were carried out by Fouriertransform
infrared spectroscopy (FT-IR) and High-performance liquid chromatography (HPLC).
Total contents of polysaccharides, beta-glucans, phenolic compounds, and flavonoids were investigated
utilizing the phenol-sulfuric acid method, enzymatic-based commercial test kit, Folin-Ciocalteu
method, and aluminium chloride colorimetric method, respectively. Antioxidant activity was determined
by using 2,2-diphenyl-1-picrylhydrazyl-hydrate (DPPH) radical scavenging assay and 2,2-
azino-bis (3-ethylbenzothiazol-6-sulfonic acid) (ABTS) radical cation decolorization assay.

Results:
Optimal conditions for the production of PEPRM included a ratio of 51.29 mL water to 1 g
PR mycelia and an extraction time of 46.23 minutes, resulting in a total polysaccharide content of
577.5 mg/g of PEPRM. FT-IR spectra of PEPRM showed two broad bands at 3272.08 cm-1 and
2924.8 cm-1 in the carbohydrate region and the peaks at 1078.44, 1019.05, and 853.0 cm-1 indicated
the presence of the pyranose ring skeleton, glycosidic linkage, and glucans. PEPRM had molar ratios
of glucose: mannose: rhamnose: fucose, i.e., 21.86: 1.00: 2.08: 3.40, respectively. PEPRM had total
contents of beta-glucans, phenolic compounds, and flavonoids as percentages of dry weight, i.e.,
21.22, 2.51, and 5.71, respectively. PEPRM showed better inhibitory activity against ABTS radicals
than DPPH radicals.

result:
Optimal conditions for the production of PEPRM were a ratio of 51.29 ml water to 1 g PR mycelia and extraction time of 46.23 min, yielding a total polysaccharide content of 577.5 mg/g of PEPRM. FT-IR spectra of PEPRM showed two broad bands at 3272.08 cm-1 and 2924.8 cm-1 in the carbohydrate region and the peaks at 1078.44, 1019.05, and 853.0 cm-1 indicated the presence of the pyranose ring skeleton, glycosidic linkage, and glucans. PEPRM had molar ratios of glucose: mannose: rhamnose: fucose, i.e., 21.86: 1.00: 2.08: 3.40, respectively. PEPRM had total contents of beta-glucan, phenolic compounds, and flavonoids as percentages of dry weight, i.e., 21.22, 2.51, and 5.71, respectively. PEPRM showed better inhibitory activity against ABTS radicals than DPPH radicals.

Conclusion:
This is the first finding to reveal that ultrasonic-assisted extraction with RSM was an
environmentally friendly alternative to produce antioxidant polysaccharides from cultured PR mycelia.

conclusion:
This is the first finding to reveal that ultrasonic-assisted extraction with RSM was an environmentally friendly alternative to produce antioxidant polysaccharides from cultured PR mycelia.