Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Enhanced Production of Exopolysaccharides by Fed-batch Culture of Ganoderma resinaceum DG-6556

  • Kim Hyun-Mi (Department of Biotechnology, College of Engineering, Daegu University) ;
  • Paik Soon-Young (Department of Microbiology, College of Medicine, The Catholic University of Korea) ;
  • Ra Kyung-Soo (Department of Food and Nutrition, Daegu Technical College) ;
  • Koo Kwang-Bon (Department of Bioindustry, College of Life & Environment, Daegu University) ;
  • Yun Jong-Won (Department of Biotechnology, College of Engineering, Daegu University) ;
  • Choi Jang-Won (Department of Bioindustry, College of Life & Environment, Daegu University)
  • Published : 2006.04.01
Download PDF
⟨ Previous Next ⟩

Abstract

The objectives of this study were to optimize submerged culture conditions of a new fungal isolate, Ganorderma resinaceum, and to enhance the production of bioactive mycelial biomass and exopolysaccharides (EPS) by fed-batch culture. The maximum mycelial growth and EPS production in batch culture were achieved in a medium containing 10 g/l glucose, 8 g/l soy peptone, and 5 mM $MnCl_2$ at an initial pH 6.0 and temperature $31^{\circ}C$. After optimization of culture medium and environmental conditions in batch cultures, a fed-batch culture strategy was employed to enhance production of mycelial biomass and EPS. Five different EPS with molecular weights ranging from 53,000 to 5,257,000 g/mole were obtained from either top or bottom fractions of ethanol precipitate of culture filtrate. A fed-batch culture of G. resinaceum led to enhanced production of both mycelial biomass and EPS. The maximum concentrations of mycelial biomass (42.2 g/l) and EPS (4.6 g/l) were obtained when 50 g/l of glucose was fed at day 6 into an initial 10 g/l of glucose medium. It may be worth attempting with other mushroom fermentation processes for enhanced production of mushroom polysaccharides, particularly those with industrial potential.

Keywords

References

  1. Bae, J.T., J. Sinha, J.P. Park, C.H. Song, and J.W. Yun. 2000. Optimization of submerged culture conditions for exobiopolymer production by Paecilomyces japonica. J. Microbiol. Biotechnol. 10, 482-487
  2. Bhargava, S., M.P. Nandakumar, A. Roy, K.S. Wenger, and M.R. Marten. 2002. Pulsed feeding during fed-batch fungal fermentation leads to reduced viscosity with out detrimentally affecting protein expression. Biotechnol. Bioeng. 81, 341-347
  3. Cao, Q.Z. and Z.B. Lin. 2004. Antitumor and anti-angiogenic activity of Ganoderma lucidum polysaccharides peptide. Acta. Pharmacol. Sin. 25, 833-838
  4. Carbone, I. and L.M. Kohn. 1993. Ribosomal DNA sequence divergence within internal transcribed spacer 1 of the Selerotiniaccae. Mycologia. 85, 415-427 https://doi.org/10.2307/3760703
  5. De Baets, S., S. Du Laing, C. François, and E.J. Vandamme. 2002. Optimization of exopolysaccharide production by Tremella mesenterica NRRL Y-6158 through implementation of fed-batch fermentation. J. Ind. Microbiol. Biotechnol. 29, 181-184 https://doi.org/10.1038/sj.jim.7000276
  6. Dubois, M., K.A. Gillis, J.K. Hamilton, P.A. Febers, and F. Smith. 1956. Colorimetric method for determination of sugar and related substances. Anal. Chem. 28, 350-356 https://doi.org/10.1021/ac60111a017
  7. Hwang, H.J., S.W. Kim, C.P. Xu, J.W. Choi, and J.W. Yun. 2003. Production and molecular characteristics of four groups of exopolysaccharides from submerged culture of Phellinus gilvus. J. Appl. Microbiol. 94, 708-719 https://doi.org/10.1046/j.1365-2672.2003.01903.x
  8. Jonathan, S.G. and I.O. Fasidi. 2001. Effect of carbon, nitrogen and mineral sources on growth of Psathyerella atroumbonata (Pegler), a Nigerian edible mushroom. Food Chem. 72, 479-483 https://doi.org/10.1016/S0308-8146(00)00265-X
  9. Kabir, Y., S. Kimura, and T. Tamura. 1988. Dietary effect of Ganoderma lucidum mushroom on blood pressure and lipid levels in spontaneously hypertensive rats (SHR). J. Nutr. Sci. Vitaminol. 34, 433-438 https://doi.org/10.3177/jnsv.34.433
  10. Kanmatsuse, K., N. Kajiwara, K. Hayashi, S. Shimogaichi, I. Fukinbara, H. Ishikawa, and T. Tamura. 1985. Studies on Ganoderma lucidum. I. Efficacy against hypertension and side effects. Yakugaku. Zasshi. (in Japanese) 105, 942-947 https://doi.org/10.1248/yakushi1947.105.10_942
  11. Kawagoe, M., K. Kawakami, Y. Nakamura, K. Naoe, K. Miki, and H. Noda. 1999. Submerged culture of Tricholoma matsutake mycelium in bubble column fermentors. J. Biosci. Bioeng. 87, 116-118 https://doi.org/10.1016/S1389-1723(99)80020-6
  12. Kim, S.D. and H.J. Nho. 2004. Isolation and characterization of alpha-glucosidase inhibitor from Ganoderma lucidum. J. Microbiol. 42, 223-227
  13. Kim, S.W., C.P. Xu, H.J. Hwang, J.W. Choi, C.W. Kim, and J.W. Yun. 2003. Production and characterization of exopolysaccharides from an enthomopathogenic fungus Cordyceps militaris NG3. Biotechnol. Prog. 19, 428-435 https://doi.org/10.1021/bp025644k
  14. Lakshmi, B., T.A. Ajith, N. Sheena, N. Gunapalan, and K.K. Janardhanan. 2003. Antiperoxidative, anti-inflammatory, and antimutagenic activities of ethanol extract of the mycelium of Ganoderma lucidum occurring in South India. Teratog. Carcinog. Mutagen. 23, 85-97 https://doi.org/10.1002/tcm.10065
  15. Lee, J.S., M.O. Lim, K.Y. Cho, J.H. Cho, S.Y. Chang, and D.H. Nam. 2006. Identification of medicinal mushroom species based on nuclear large subunit rDNA sequences. J. Microbiol. 44, 29-34
  16. Lee, S.Y. and H.M. Rhee. 1990. Cardiovascular effects of mycelium extract of Ganoderma lucidum: inhibition of sympathetic outflow as a mechanism of its hypotensive action. Chem. Pharm. Bull. 38, 1359-1364 https://doi.org/10.1248/cpb.38.1359
  17. Lee, K.M., S.Y. Lee, and H.Y. Lee. 1999. Bistage control of pH for improving exopolysaccharide production from mycelia of Ganoderma lucidum in an air-lift fermentor. J. Biosci. Bioeng. 88, 646-650 https://doi.org/10.1016/S1389-1723(00)87094-2
  18. Lee, J.M., H.J. Kwon, H. Jeong, J.W. Lee, S.Y. Lee, and S.J. Baek. 2001. Inhibition of lipid peroxidation and oxidative DNA damage by Ganoderma lucidum. Phytother. Res. 15, 245-249 https://doi.org/10.1002/ptr.830
  19. Lin, J.M., C.C. Lin, H.F. Chiu, and J.J. Yang. 1993. Evaluation of the anti-inflammatory and liver-protective effects of Anoectochilus formosanus, Ganoderma lucidum and Gynostemma pentaphyllum in rats. Am. J. Chin. Med. 21, 59-70 https://doi.org/10.1142/S0192415X9300008X
  20. Liu, Y.H., C.F. Tsai, M.C. Kao, Y.L. Lai, and J.J. Tsai. 2003. Effectiveness of Dp2 nasal therapy for Dp2-induced airway inflammation in mice: using oral Ganoderma lucidum as an immunomodulator. J. Microbiol. Immunol. Infect. 36, 236-242
  21. Mizuno, T., G. Wang, J. Zhang, H. Kawagish, T. Nishitoba, and J. Li. 1995. Ganoderma lucidum and Ganoderma tsugae: Bioactive substances and medicinal effects. Food Rev. Int. 11, 151-166 https://doi.org/10.1080/87559129509541025
  22. Ng, P.C., Y.C. Kong, K.M. Ko, and C.M. So. 1993. Antioxidant activity of Ganoderma lucidum: protective effects on carbon tetrachloride-induced hepatotoxicity. Acta Hortic. 332, 219-226
  23. Ooi, V.E. and F. Liu. 2000. Immunomodulation and anti-cancer activity of polysaccharide-protein complexes. Curr. Med. Chem. 7, 715-730 https://doi.org/10.2174/0929867003374705
  24. Park, J.P., S.W. Kim, H.J. Hwang, and J.W. Yun. 2001. Optimization of submerged culture conditions for the mycelial growth and exo-biopolymer production by Cordyceps militaris. Lett. Appl. Microbiol. 33, 76-81 https://doi.org/10.1046/j.1472-765X.2001.00950.x
  25. Shiao, M.S., K.R. Lee, N.L. Lee, and T.W. Chemg. 1994. Natural products and biological activities of the Chinese medicinal fungus Ganoderma lucidum. ACS. Symp. Ser. 547, 342-354
  26. Shimizu, A., T. Yano, Y. Saito, and Y. Inada. 1985. Isolation of an inhibitor of platelet aggregation from a fungus, Ganoderma lucidum. Chem. Pharm. Bull. 33, 3012-3015 https://doi.org/10.1248/cpb.33.3012
  27. Tang, Y.J. and J.J. Zhong. 2002. Fed-batch fermentation of Ganoderma lucidum for hyperproduction of polysaccharide and ganoderic acid. Enzyme Microb. Technol. 31, 20-28 https://doi.org/10.1016/S0141-0229(02)00066-2
  28. Tasaka, K., M. Akagi, K. Miylshi, M. Mio, and T. Makino. 1988. Antiallergic constituents in the culture medium of Ganoderma lucidum. I. Inhibitory effect of cyclooctasulfur on histamine release. Agents. Actions 23, 153-156 https://doi.org/10.1007/BF02142526
  29. Thompson, J.D., D.G. Higgins, and T.J. Gibson. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalies and weight matrix choice. Nucleic Acids Res. 22, 4673-4680 https://doi.org/10.1093/nar/22.22.4673
  30. Wagner, R., D.A. Mitchell, G.L. Sassaki, M.A.L.A. Amazonas, and M. Berovic. 2003. Current techniques for the cultivation of Ganoderma lucidum for the production of biomass, ganoderic acid and polysaccharides. Food Technol. Biotechnol. 41, 371-382
  31. Wagner, R., D.A. Mitchell, G.L. Sassaki, and M.A.L.A. Amazonas. 2004. Links between morphology and physiology of Ganoderma lucidum in submerged culture for the production of exopolysaccharide. J. Biotechnol. 114, 153-164 https://doi.org/10.1016/j.jbiotec.2004.06.013
  32. Waytt, P.J. 1993. Light scattering and the absolute characterization of macromolecules. Anal. Chim. Acta 272, 1-40 https://doi.org/10.1016/0003-2670(93)80373-S
  33. Wu, T.S., L.S. Shi, and S.C. Kuo. 2001. Cytotoxicity of Ganoderma lucidum Triterpenes. J. Nat. Prod. 64, 1121-1122 https://doi.org/10.1021/np010115w
  34. Yang, B.K., S.C. Jeong, and C.H. Song. 2002. Hypolipidemic effect of exo- and endo-biopolymers produced from submerged mycelial culture of Ganoderma lucidum in rats. J. Microbiol. Biotechnol. 12, 872-877
  35. Yang, B.K., M.A. Wilson, K.Y. Cho, and C.H. Song. 2004. Hypoglycemic effect of exo- and endo-biopolymers produced by submerged mycelial culture of Ganoderma lucidum in streptozotocin-induced diabetic rats. J. Microbiol. Biotechnol. 14, 972-977
  36. Zhang, H.N. and Z.B. Lin. 2004. Hypoglycemic effect of Ganoderma lucidum polysaccharides. Acta. Pharmacol. Sin. 25, 191-195