Effect of deproteinization methods on the antioxidant activity of polysaccharides extracted from Lentinus edodes stipe

  • Shunfeng LiEmail author
  • Anjian Wang
  • Lina Liu
  • Guangrui Tian
  • Fangfang Xu
Original Paper


Removal of protein from plant polysaccharide extract is often regarded as essential step for antioxidant activity evaluation. The aim of this study was to compare two different methods (neutral protease and Sevag) for efficient removal of protein from Lentinus edodes stipe polysaccharide extract. Orthogonal test was used to optimize the parameters of neutral protease deproteinization effects on polysaccharide extracted from Lentinus edodes stipe. Furthermore, antioxidant activity was also investigated by the two deproteinization methods. Results showed that the optimum parameters of deproteinization by neutral protease were pH of 6, temperature of 60 °C, time of 1 h, and enzyme amount 0.2%. Under these conditions, the polysaccharide retention rate was 92.29%, and the deproteinization rate was 84.30%. This result was higher than that of Sevag, which deproteinization rate and polysaccharide retention rate were 55.43% and 82.29%, respectively. The antioxidant activity of polysaccharide declined after the polysaccharide deproteinized for both techniques, however, the polysaccharide deproteinized by Sevag showed the lowest antioxidant activity. Therefore, neutral protease could be used for deproteinization of polysaccharide extracted from Lentinus edodes stipe.


Lentinus edodes stipe Polysaccharide Deproteinization Neutral protease Sevag Antioxidant activity 



This work was financially supported by the Special Fund for Independent Innovation Research (Grant No. YUCAIKE [2018]9) established by Henan Academy of Agricultural Sciences, Scientific and Technological Project of Henan Province (Grant Nos. 182102110276, 172102110085), China.


  1. 1.
    A. Andō, On the components of pileus and stipe of dried shiitake (Lentinus edodes (Berk.) Sing): chemical components and hot water soluble polysaccharides. J. Home Econ. Jpn. 27(2), 86–91 (1976)Google Scholar
  2. 2.
    P.S. Bisen, R.K. Baghel, B.S. Sanodiya, G.S. Thakur, G.B.K. Prasad S, Lentinus edodes: a macrofungus with pharmacological activities. Curr. Med. Chem. 17(22), 2419–2430 (2010)CrossRefGoogle Scholar
  3. 3.
    M.M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72(1–2), 248–254 (1976)CrossRefGoogle Scholar
  4. 4.
    W. Chen, W. Li, Y. Yang, H. Yu, S. Zhou, J. Feng, X. Li, Y. Liu, Analysis and evaluation of tasty components in the pileus and stipe of Lentinula edodes at different growth stages. J. Agric. Food Chem. 63(3), 795–801 (2015)CrossRefGoogle Scholar
  5. 5.
    China Edible Fungi Association. (2017). Notice on the results of 2016 annual output, output value and export statistics of edible fungus in China (China Edible Fungi Association [2017]11). Beijing. Available from Accessed 24 Oct 2017
  6. 6.
    Y. Chi, X. Wang, R. Wang, Z. Wang, Y. Wu, H. Jiang, F. Zhou, C. Hua, Comparative investigation into deproteinization methods for Cordyceps militaris polysaccharides. Food Sci. Technol. 39(12), 202–206 (2014)Google Scholar
  7. 7.
    M. Darré, L. Valerga, L.C. Ortiz Araque, M.L. Lemoine, P.V. Demkura, A.R. Vicente, A. Concellón, Role of UV-B irradiation dose and intensity on color retention and antioxidant elicitation in broccoli florets (Brassica oleracea var. Italica). Postharvest Biol. Technol. 128, 76–82 (2017)CrossRefGoogle Scholar
  8. 8.
    M. Dermiki, N. Phanphensophon, D.S. Mottram, L. Methven, Contributions of non-volatile and volatile compounds to the umami taste and overall flavour of shiitake mushroom extracts and their application as flavour enhancers in cooked minced meat. Food Chem. 141(1), 77–83 (2013)CrossRefGoogle Scholar
  9. 9.
    M. Dubois, K.A. Gilles, J.K. Hamilton, P.A. Rebers, F. Smith, Colorimetric method for determination of sugars and related substances. Anal. Chem. 28(3), 350–356 (1956)CrossRefGoogle Scholar
  10. 10.
    H. Gao, Y. Wang, D. Shi, S. Wang, J. He, J. Guan, Radio-frequency assisted extraction for polysaccharides from stems of shiitake mushroom. Trans. Chin. Soc. Agric. Eng. 25(S1), 180–184 (2009)Google Scholar
  11. 11.
    Y. Jiang, On the technology of ultrasonic-assisted polysaccharide extraction of shii-take stipe. Value Eng. (3), 137–139 (2016)Google Scholar
  12. 12.
    S. Li, A. Wang, L. Liu, G. Tian, S. Wei, F. Xu, Evaluation of nutritional values of shiitake mushroom (Lentinus edodes) stipes. J. Food Meas. Charact. 12(3), 2012–2019 (2018)CrossRefGoogle Scholar
  13. 13.
    S. Li, A. Wang, L. Liu, G. Tian, F. Xu (2018). Extraction of polysaccharides under vacuum condition from Lentinus edodes stipe and their antioxidant activities in vitro. Food Sci. Biotechnol. Google Scholar
  14. 14.
    S. Li, L. Zhang, J. Fu, X. Liu, Antioxidant properties of polysaccharide extracts from fruitbodies of Hypsizigus marmoreus. Acta Agric. Boreali-occidentalis Sinica 17(4), 302–305 (2008)Google Scholar
  15. 15.
    L.-Y. Lin, Y.-H. Tseng, R.-C. Li, J.-L. Mau, Quality of shiitake stipe bread. J. Food Process. Preserv. 32(6), 1002–1015 (2008)CrossRefGoogle Scholar
  16. 16.
    P.-H. Lin, S.-Y. Huang, J.-L. Mau, B.-K. Liou, T.J. Fang, A novel alcoholic beverage developed from shiitake stipe extract and cane sugar with various Saccharomyces strains. LWT—Food Sci. Technol. 43(6), 971–976 (2010)Google Scholar
  17. 17.
    N. Qin, W.-Y. Miao, W.-Q. Lian, Y.-L. Sun, Optimization of microwave-assisted extraction of polysaccharides from Lentinus edodes root by response surface analysis and its antioxidant activity. J. Food Saf. Qual. 8(2), 594–600 (2017)Google Scholar
  18. 18.
    T. Rahman, M. Choudhury, Shiitake mushroom: a tool of medicine. Bangladesh J. Med. Biochem. 5(1), 24–32 (2012)CrossRefGoogle Scholar
  19. 19.
    D. Shi, H. Gao, M. Zhou, W. Cheng, S. Wang, X. Du, D. Yang, Effect of super fine crushing on the extraction yield of polysaccharide in Lentinus edodes stem. Hubei Agric. Sci. 48(7), 1730–1732 (2009)Google Scholar
  20. 20.
    Q. Shi, W. Shao, Determination of nutritive components of eight edible fungi. J. Gansu Agric. Univ. 38(3), 223–339 (2003)Google Scholar
  21. 21.
    Y. Tian, H. Zeng, Z. Xu, B. Zheng, Y. Lin, C. Gan, Y.M. Lo, Ultrasonic-assisted extraction and antioxidant activity of polysaccharides recovered from white button mushroom (Agaricus bisporus). Carbohyd. Polym. 88(2), 522–529 (2012)CrossRefGoogle Scholar
  22. 22.
    S.-Y. Tsai, S.-J. Huang, J.-L. Mau, Antioxidant properties of hot water extracts from Agrocybe cylindracea. Food Chem. 98(4), 670–677 (2006)CrossRefGoogle Scholar
  23. 23.
    A. Wang, J. Li, X. Wang, S. Li, L. Liu, G. Tian, Decolorization of dietary fiber from fermented Lentinus edodes stipe by the combination of ultrasound and H2O2. Packag. Food Mach. 35(1), 19–23 (2017)Google Scholar
  24. 24.
    J.-H. Yang, H.-C. Lin, J.-L. Mau, Non-volatile taste components of several commercial mushrooms. Food Chem. 72(4), 465–471 (2001)CrossRefGoogle Scholar
  25. 25.
    M.-T. Yen, J.-L. Mau, Preparation of fungal chitin and chitosan from shiitake stipes. Fungal Sci. 21(1–2), 1–11 (2006)Google Scholar
  26. 26.
    M.-T. Yen, J.-L. Mau, Selected physical properties of chitin prepared from shiitake stipes. LWT—Food Sci. Technol. 40(3), 558–563 (2007)Google Scholar
  27. 27.
    Z. Zhang, H. Song, Z. Peng, Q. Luo, J. Ming, G. Zhao, Characterization of stipe and cap powders of mushroom (Lentinus edodes) prepared by different grinding methods. J. Food Eng. 109(3), 406–413 (2012)CrossRefGoogle Scholar
  28. 28.
    J. Zheng, X. Ding, Study on the antimutative PJF1 polysaccharide from foot body of Lentinus edodes. J. Wuxi Univ. Light Ind. 14(1), 7–13 (1995)Google Scholar
  29. 29.
    S. Zheng, Y. Peng, Q. Chen, M. Zheng, Biological activity of Tremella polysaccharide with or without deproteinization. Wuyi Sci. J. 33(1), 94–100 (2017)Google Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Agro-Products ProcessingHenan Academy of Agricultural SciencesZhengzhouPeople’s Republic of China

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