Production of Lignin Peroxidase by Phellinus igniarius and Cytotoxic Effects of Lignin Hydrolysates Derived from Wood Biomass on Cancer Cells

  • Published : 2004.09.01

Abstract

Over the past several years, research efforts have been directed both at economically producing valuable substances from the wood biomass and at producing lignolytic enzymes at a lower cost. In the present study, we found that Phellinus igniarius, the basidiomycetes, secreted lignin peroxidase as a main lignolytic enzyme, which was detected maximum activity at 16th day of culture and showed 37 kDa of molecular mass in identification by activity assay and purification by anion-exchange chromatography. The Phellinus igniarius-derived lignin peroxidase hydrolyzed steam-exploded wood (Quercus mongolica) powder into small molecules showing cytotoxicity against cancer cel1s (HepG2 hepatoma, SK-N-SH neuroblastoma, B16 melanoma, MBT-2 bladder cancer). In addition, the enzyme hydrlysates of lignins (ELg) that were extracted from the steam-exploded oak showed more potent cytotoxic effects on the cancer cells than the enzyme hydrolysates of wood biomass (EWp), indicating that the cytotoxic effect of EWp may be due to the enzyme-degraded products of lignin among the lignocellulosics. Furthermore, the cytotoxic effect of ELg on Chang, normal liver cells, was much less potent than that of ELg on HepG2 and B16 cancer cells, indicating that the cytotoxic effect of ELg may be specific for cancer cells. The present results suggest that Phellinus igniarius may be a useful resource for the large-scale production of lignin peroxidase and that the lignin peroxidase may be applied for the generation of valuable biodegradation products from wood lignocellulosics for medical use.

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References

  1. Eriksson, K. E. (1993) Concluding remarks: where do we stand and where are we going? Lignin biodegradation and practical utilization. J. Biotechnol. 30, 149-158 https://doi.org/10.1016/0168-1656(93)90036-M
  2. Field, J. A., de Jong, E., Feijoo Costa, G., de Bont, J. A. (1992) Biodegradation of polycyclic aromatic hydrocarbons by new isolates of white rot fungi. Appl Environ Microbiol. 58, 2219-2226
  3. Gordaliza, M., Castro, M. A, del Corral, J. M., Feliciano, A. S. (2000) Antitumor properties of podophyllotoxin and related compounds. Curr Pharm Des. 6, 1811-1839 https://doi.org/10.2174/1381612003398582
  4. Gordaliza, M., Miguel del Corral, J. M., Angeles Castro, M., Garcia- Garcia, P. A., San Feliciano, A. (2001) Cytotoxic cyclolignans related to podophyllotoxin. Farmaco. 56, 297-304 https://doi.org/10.1016/S0014-827X(01)01030-8
  5. Ichimura, T, Otake, T., Mori, H., Maruyama, S. (1999) HIV-1 protease inhibition and anti-HIV effect of natural and synthetic water-soluble lignin-like substances. Biosci. Biotechnol. Biochem. 63, 2202-2204 https://doi.org/10.1271/bbb.63.2202
  6. Kirk, T. K. and Farrell, R. L. (1987) Enzymatic 'combustion'; the microbial degradation of lignin, Annu. Rev. Microbial. 41, 465-505 https://doi.org/10.1146/annurev.mi.41.100187.002341
  7. Kofujita, H., Asad, Y, Kuwahara, M. (1991) Alkyl-aryl cleavage of phenolic $\beta$ -o-4 lignin substructure model compound by Mn (Ⅱ)-peroxidase isolated from Pleurotus ostreatus. Mokuzai Gakkaishi 37, 555-561
  8. Lee, J. (1997) Biological conversion of lignocellulosic biomass to ethanol. J Biotechnol. 56, 1-24 https://doi.org/10.1016/S0168-1656(97)00073-4
  9. Mikulasova, M. and Kosikova, B. (2003) Modulation of mutagenicity of various mutagens by lignin derivatives. Mutat. Res. 535, 171-180 https://doi.org/10.1016/S1383-5718(02)00319-4
  10. Pelaez, F, Martinez, M. J., Martinez, A. T. (1995) Screening of 68 species of basidiomycetes for enzymes involved in lignin degradation. Mycol. Res. 99, 37-42 https://doi.org/10.1016/S0953-7562(09)80313-4
  11. Sakagami, H., Takeda, M., Kawazoe, Y., Nagata, K., Ishihama, A., Ueda, M., Yamazaki, S. (1992) Anti-influenza virus activity of a lignin fraction from cone of Pinus parviflora Sieb. et Zucc. In Vivo 6, 491-495
  12. Sugiura, M., Hirai, H., Nishida, T. (2003) Purification and characterization of a novel lignin peroxidase from white-rot fungus Phanerochaete sordida YK-624. FEMS Microbiol Lett. 224, 285-290 https://doi.org/10.1016/S0378-1097(03)00447-6
  13. Tien, M. and Kirk, T. K. (1983) Lignin-degrading enzyme from the hymenomycete Phanerocheate chrysosporium burds. Science 221, 661-663 https://doi.org/10.1126/science.221.4611.661
  14. Tien, M. and Kirk, T. K. (1984) Lignin degrading enzyme from Phanerocheate chrysosporium: purification, characterization and catalytic properties of a unique $H^2O^2$ -requiring oxygenase. Proc. Natl. Acid. Sci. USA 81, 2280-2284
  15. van de Loosdrecht, A. A., Nennie, E., Ossenkoppele, G. J., Beelen, R. H., Langenhuijsen, M. M. (1991) Cell mediated cytotoxicity against U 937 cells by human monocytes and macrophages in a modified colorimetric MTT assay. A methodological study. J. Immunol. Methods 141, 15-22. https://doi.org/10.1016/0022-1759(91)90205-T