Abstract: An ultrasonic-assisted technique was employed to extract crude polysaccharide from Tricholoma matsutake fruiting bodies. Single-factor tests and orthogonal experimental design (L₉(3³)) were used to obtain the optimal extraction conditions. Results showed that the optimal parameters were as follows: ultrasonic temperature, 40 °C; ultrasonic time, 50 min; water to raw material ratio, 25 ml/g; ultrasonic frequency, 45 kHz; and ultrasonic power, 100 W. Three novel T. matsutake polysaccharide (TMP) fractions (TMP30, TMP60, and TMP80) were isolated and purified from TMP by stepwise alcohol precipitation. Their preliminary structural features were determined by high-performance anion-exchange chromatography with pulsed-amperometric detection (HPAEC-PAD) and Fourier transform infrared spectrophotometer (FT-IR) analyses. Furthermore, their in vitro antioxidant activity was investigated in terms of a reducing power assay and the scavenging rates of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radicals. The order of the various fractions based on their antioxidant activity was TMP80>TMP>TMP60>TMP30. These findings suggested that novel polysaccharide fractions from T. matsutake, especially TMP80, could be promising active macromolecules for biomedical use.

松茸多糖的超声波提取优化、结构特性和抗氧化活性研究

[1]Achat, S., Tomao, V., Madani, K., et al., 2012. Direct enrichment of olive oil in oleuropein by ultrasound-assisted maceration at laboratory and pilot plant scale. Ultrason. Sonochem., 19(4):777-786.

[2]Afshari, K., Samavati, V., Shahidi, S.A., 2015. Ultrasonic-assisted extraction and in-vitro antioxidant activity of polysaccharide from Hibiscus leaf. Int. J. Biol. Macromol., 74:558-567.

[3]Ardestani, A., Yazdanparast, R., 2007. Antioxidant and free radical scavenging potential of Achillea santolina extracts. Food Chem., 104(1):21-29.

[4]Bohn, J.A., BeMiller, J.N., 1995. (1→3)-β-D-Glucans as biological response modifiers: a review of structure-functional activity relationships. Carbohydr. Polym., 28(1):3-14.

[5]Brand-Williams, W., Cuvelier, M.E., Berset, C., 1995. Use of a free radical method to evaluate antioxidant activity. LWT-Food Sci. Technol., 28(1):25-30.

[6]Chemat, F., Rombaut, N., Sicaire, A., et al., 2017. Ultrasound assisted extraction of food and natural products. Mechanisms, techniques, combinations, protocols and applications. A review. Ultrason. Sonochem., 34:540-560.

[7]Devasagayam, T.P.A., Tilak, J.C., Boloor, K.K., et al., 2004. Free radicals and antioxidants in human health: current status and future prospects. J. Assoc. Phys. India, 52:794-804.

[8]Du, X.J., Yang, Y., Ye, L.B., et al., 2009. Structural elucidation and immuno-stimulating property of an acidic heteropolysaccharide (TAPA1) from Tremella aurantialba. Carbohydr. Res., 344(5):672-678.

[9]Du, X.J., Mu, H.M., Zhou, S., et al., 2013. Chemical analysis and antioxidant activity of polysaccharides extracted from Inonotus obliquus sclerotia. Int. J. Biol. Macromol., 62(11):691-696.

[10]Hoshi, H., Iijima, B., Ishihara, Y., et al., 2008. Absorption and tissue distribution of an immunomodulatory α-D-glucan after oral administration of Tricholoma matsutake. J. Agric. Food Chem., 56(17):7715-7720.

[11]Hu, J., Jia, X.J., Fang, X.B., et al., 2016. Ultrasonic extraction, antioxidant and anticancer activities of nove lpolysaccharides from Chuanxiong rhizome. Int. J. Biol. Macromol., 85:277-284.

[12]Huang, S.Q., Ning, Z.X., 2010. Extraction of polysaccharide from Ganoderma lucidum and its immune enhancement activity. Int. J. Biol. Macromol., 47(3):336-341.

[13]Kim, S.S., Lee, J.S., Cho, J.Y., et al., 2010a. Effects of C/N ratio and trace elements on mycelial growth and exo-polysaccharide production of Tricholoma matsutake. Biotechnol. Bioproc. E, 15(2):293-298.

[14]Kim, S.S., Lee, J.S., Cho, J.Y., et al., 2010b. Process development for mycelial growth and polysaccharide production in Tricholoma matsutake liquid culture. J. Biosci. Bioeng., 109(4):351-355.

[15]Li, Y., Fabiano-Tixier, A.S., Tomao, V., et al., 2013. Green ultrasound-assisted extraction of carotenoids based on the bio-refinery concept using sunflower oil as an alternative solvent. Ultrason. Sonochem., 20(1):12-18.

[16]Luo, Q., Zhang, J., Yan, L., et al., 2011. Composition and antioxidant activity of water-soluble polysaccharides from Tuber indicum. J. Med. Food., 14(12):1609-1616.

[17]Lv, Y., Yang, X.B., Zhao, Y., et al., 2009. Separation and quantification of component monosaccharides of the tea polysaccharides from Gynostemma pentaphyllum by HPLC with indirect UV detection. Food Chem., 112(3):742-746.

[18]Murphey, J.M., Spayd, S.E., Powers, J.R., 1989. Effect of grape maturation on soluble protein characteristics of Gewurztraminer and White Riesling juice and wine. Am. J. Enol. Viticult., 40(3):199-207.

[19]Pingret, D., Durand, G., Fabiano-Tixier, A., et al., 2012. Degradation of edible oil during food processing by ultrasound: electron paramagnetic resonance, physicochemical, and sensory appreciation. J. Agric. Food Chem., 60(31):7761-7768.

[20]Smironff, N., Cumbes, Q.J., 1989. Hyroxyl radical scavenging activity of compatible solutes. Phytochemistry, 28(4):1051-1060.

[21]Tursun, K., Zhan, R., Zhang, H., et al., 2010. Study on antioxidant activity of Acroptilon repens. Lett. Biol., 21(3):406-411 (in Chinese).

[22]Wang, N.N., Zhang, Y., Wang, X.P., et al., 2016. Antioxidant property of water-soluble polysaccharides from Poria cocos Wolf using different extraction methods. Int. J. Biol. Macromol., 83:103-110.

[23]Wang, Y.F., Yang, Z.W., Wei, X.L., 2012. Antioxidant activities potential of tea polysaccharide fractions obtained by ultrafiltration. Int. J. Biol. Macromol., 50(3):558-564.

[24]Xu, F., Jabasini, M., Baba, Y., 2005. Screening of mixed poly (ethylene oxide) solutions for microchip separation of double-stranded DNA using an orthogonal design approach. Electrophoresis, 26(15):3013-3020.

[25]Yang, S., Ren, X.D., Sheng, J.X., et al., 2010. Preparation and the antitumor activity in vitro of polysaccharides from Tricholoma matsutake. W. J. Microbiol. Biotechnol., 26(3):497-503.

[26]Ye, C., Han, N., Teng, F.K., et al., 2013. Extraction optimization of polysaccharides of Schisandrae Fructus and evaluation of their analgesic activity. Int. J. Biol. Macromol., 57(6):291-296.

[27]Ye, L.B., Zhang, J.S., Yang, Y., et al., 2009. Structural characterisation of a heteropolysaccharide by NMR spectra. Food Chem., 112(4):962-966.

[28]Yin, X.L., You, Q.H., Jiang, Z.H., 2009. Extraction and purification of Tricholoma matsutake polysaccharides. China Brewing, 211(10):171-173.

[29]Yin, X.L., You, Q.H., Jiang, Z.H., 2011. Optimization of enzyme assisted extraction of polysaccharides from Tricholoma matsutake by response surface methodology. Carbohydr. Polym., 86(3):1358-1364.

[30]You, L.J., Gao, Q., Feng, M.Y., et al., 2013. Structural characterisation of polysaccharides from Tricholoma matsutake and their antioxidant and antitumour activities. Food Chem., 138(4):2242-2249.

[31]You, Q.H., Yin, X.L., Ji, C.W., 2014. Pulsed counter-current ultrasound-assisted extraction and characterization of polysaccharides from Boletus edulis. Carbohydr. Polym., 101(1):379-385.

[32]Zhang, W.J., 1999. Biochemical Research Technology on Glycoconjugates, 2nd Ed. Zhejiang University Press, Hangzhou (in Chinese).

[33]Zhao, Z., Xu, X., Ye, Q., et al., 2013. Ultrasound extraction optimization of Acanthopanax senticosus polysaccharides and its antioxidant activity. Int. J. Biol. Macromol., 59(4):290-294.