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Ultrasound-Assisted Enzymatic Extraction of Polysaccharides from Tricholoma matsutake: Optimization, Structural
Wen Gao1, Yang Wang1, Fuping Lu1
1Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China.
Abstract:
This study optimized ultrasound-assisted enzymatic (UAE) extraction of TMP (Tricholoma matsutake polysaccharide) through response surface methodology. The optimal conditions included complex enzyme comprising 1.15% cellulase, 0.60% pectinase, and 0.95% dispase, with ultrasound for 24 min at 84.5 °C and enzyme hydrolysis at pH 5.0. This process yielded 19.74 ± 0.51% TMP, exceeding traditional hot water extraction by over four times. Fourier transform infrared spectroscopy (FT-IR) confirmed that UAE did not alter the structure of TMP. In vitro experiments indicated that TMP-UAE demonstrated enhanced antioxidant properties. Further purification through DEAE-52 and Sephadex G-100 chromatography resulted in a homogenous polysaccharide fraction (TMP). Characterization indicated that TMP has an average molecular weight of 2.79 × 104 Da, composed of fucose, galactose, glucose and mannose in a 2.00:9.44:86.29:2.28 molar ratio. FT-IR indicated the presence of C-O-C glycosidic bonds and pyranyl-type sugar rings. Scanning electron microscopy displayed loose lamellar structures with small pores. Finally, TMP exhibited therapeutic potential against C. elegans in Parkinson's disease, including reducing α-synuclein aggregation, protecting dopaminergic neurons, and prolonging lifespan. This study provides an efficient extraction method for TMP and an insight into its neuroprotective effect in PD C.elegans.
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