Production and Characterization of Manganese Peroxidase from the White Rot Fungus Pleurotus ostreatus in Liquid Culture

액체배양한 느타리 버섯균(Pleurotus ostreatus)으로부터 망간퍼옥시데이즈의 생산 및 특성

  • Lee, Jae-Sung (Department of Food Science and Technology, Yeungnam University) ;
  • Ha, Hyo-Cheol (Institute of Food & Culture, Pulmuone Co. Ltd.)
  • 이재성 (영남대학교 자연자원대학 생물자원공학부 식품가공학과) ;
  • 하효철 (풀무원 식문화 연구원)
  • Published : 2004.03.31

Abstract

The ligninolytic basidiomycete, Pleurotus ostreatus K-2946, was produced a manganese peroxidase (MnP) activity when grown in liquid culture with glucose-yeast-peptone (G-Y-P) medium. However, lignin peroxidase (LiP) was not detected in this culture medium. The purification progress of MnP was purified that included chromatography on Sepharose CL-6B, Superdex 75 prep grade and Mono-Q. MnP purified by column chromatography, was 36400 dalton and a pI of 3.95. The optimal pH and temperature of the purified MnP activity were 5.0 and $55^{\circ}C$. The characteristics of MnP produced was quite similar to those of MnP 3 isoenzyme produced by other strains of P. ostreatus.

리그닌 분해균인 느타리 버섯균(Pleurotus ostreatus) K-2946을 glucose-peptone-yeast(G-P-Y) 액체배지에서 배양 하여 망간퍼옥시데이즈를 생산하였다. 본 실험조건 하에서 리그닌 퍼옥시데이즈는 생산되지 않았다. 느타리버섯균 K-2946이 생산하는 망간 퍼옥시데이즈를 이온크로마토그래피, 겔크로마토그래피를 이용하여 정제하였다. 정제한 망간 퍼옥시데이즈의 분자량은 36400 Da이였고 pI는 3.95였다. 정제된 효소의 최적 pH는 5.0이었으며 최적온도는 $55^{\circ}C$이었다.

Keywords

References

  1. Hammel K. E., Jensen, K. A., Mozuch, M. D., Landucci, L. L., Tien, M. and Pease, E. A. (1993) Ligninolysis by a purified lignin peroxidase. J. Biotl. Chem. 268, 12274-12281
  2. Hammel K. E., Jensen, K. A., Mozuch, M. D., Landucci, L. L., Tien, M. and Pease, E. A. (1993) Ligninolysis by a purified lignin peroxidase. J. Biotl. Chem. 268, 12274-12281
  3. Wariishi, H., Valli, K., Renganathan, V. and Gold, M. H. (1989) Thiol-mediated oxidation of nonphenolic lignin model compounds by manganese peroxidase of Phanerochaete chrysosporium. J. Biol. Chem. 264, 14185-14191
  4. Arora, D. S.. Chander, M. and Gill, P. K. (2002) Involvement of lignin peroxidase, manganese peroxidase and laccase in degradation and selective ligninolysis of wheat straw. Int. Biodeter. Biodegr. 50, 115-120 https://doi.org/10.1016/S0964-8305(02)00064-1
  5. Rothschild, N., Novotny, C., Sasek, V, and Dosoretz, C. G. (2002) Ligninolytic enzymes of the fungus Irpex lacteus (Polyporus tulipiferae): isolation and characterization of lignin peroxidase. Enzyme Microb. Tech. 31, 627-633 https://doi.org/10.1016/S0141-0229(02)00171-0
  6. Martinez, A. T. (2002) Molecular biology and structure-function of lignin-degrading heme peroxidases. Enzyme Microb. Tech. 30, 425-444 https://doi.org/10.1016/S0141-0229(01)00521-X
  7. Mester, T. and Field, J. A. (1998) Characterization of a novel manganese peroxidase-lignin peroxidase hybrid isozyme produced by Bjerkandera species strain B0S55 in the absence of manganese. J. Biol Chem. 273, 15412-15417 https://doi.org/10.1074/jbc.273.25.15412
  8. Hatakka, A. (1994) Lignin-modifying enzymes from selected white-rot-fungi production and role in lignin degradation. FEMS Microbiol. Rev. 13, 125-135 https://doi.org/10.1111/j.1574-6976.1994.tb00039.x
  9. Nagarathnamma, R., Bajpai, P. and Bajpai, P. K. (1999) Studies on decolourization, degradation and detoxification of chlorinated lignin compounds in kraft bleaching effluents by Ceriporiopsis subvermispora. Process Biochem. 34, 939-948 https://doi.org/10.1016/S0032-9592(99)00021-7
  10. Reddy, C. A. (1995) The potential for white-rot fungi in the treatment of pollutants. Curr. Opin. Biotechnol. 6, 320-328 https://doi.org/10.1016/0958-1669(95)80054-9
  11. Mester, T. and Tien, M. (2000) Oxidation mechanism of ligninolytic enzymes involved in the degradation of environmental pollutants. Int. Biodeter. Biodegr. 46, 51-59 https://doi.org/10.1016/S0964-8305(00)00071-8
  12. Heinfling, A., Martinez, M. J., Martinez, A. T., Bergbauer, M. and Szewzyk, U. (1998) Purification and characterization of peroxidases from the dye-decolorizing fungus Bjerkandera adusta. FEMS Microbiol Lett. 165, 43-50 https://doi.org/10.1111/j.1574-6968.1998.tb13125.x
  13. Tien, M. and Kirk, T. K. (1983) Lignin-degrading enzyme from the hymenonycete Phanerochaete chrysosporium. Science 221, 660-661
  14. Gold M. H. and Alic, M. (1993) Molecular biology of the lignindegrading basidiomycete Phanerochaete chrysosporiwn. Microbiol. Rev. 57, 605-622
  15. Srinivasan, C, D'Souza, T. M., Boominathan, K. and Reddy, C. A. (1995) Demonstration of laccase in the white rot basidiomycete Phanerochaete. chrysosporium BKM-F1767. Appl. Environ. Microbiol. 61, 4274-4277
  16. Seo, Y. S., Ryu, W. R., Chang, Y. K. and Cho. M. H. (2000) Biodegradation of polycyclic aromatic hydrocarbons using immobilized cells of Phanerochaete chrysosporium. Kor. J. Biotech. Bioeng. 15, 247-253
  17. Sang, B. I., Kim, Y. H. and Yoo, Y. J. (1995) Induction and stabilization of lignin peroxidase from Phanerochaete chrysosporium. J. Micmbiol. Biotechnol. 5, 218-223
  18. Ryu, W. R., Shim, S. H., Jang, M. Y,, Jeon, Y, J., Oh, K. K. and Cho, M. H. (2000) Biodegradation of pentachlorophenol by white rot fungi under ligninolytic and nonligninolytic conditions. Biotechnol. Bioprocess Eng. 5, 211-214 https://doi.org/10.1007/BF02936597
  19. Kuwahara, M., Glenn, J. K., Morgan, M. A. and Gold, M. H. (1984) Separation and characterization of two extracellular $H_2O_2$ -dependent oxidase from lignmolytic cultures of Phanerochaete chrysosporium. FEBS Lett. 169, 247-250 https://doi.org/10.1016/0014-5793(84)80327-0
  20. Hofrichter, M. (2002) Review: lignin conversion by manganese peroxidase (MnP). Enz. Microb. Tech. 30, 454-466 https://doi.org/10.1016/S0141-0229(01)00528-2
  21. Tien, M and Kirk, T. K. (1984) Lignin degrading enzyme from Phanerochaete chi-ysosporium: purification, characterization and catalytic properties of a unique $H_2O_2$ -requihng oxygenase. Proc. Natl. Acad. Sci. USA 81, 2280-2284 https://doi.org/10.1073/pnas.81.8.2280
  22. Kofujita, H., Asada, Y. and 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
  23. Bradford M. M. (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal. Biochem. 11, 248-254
  24. Yoshida, S., Yonehara, S., Minami, S., Ha, H.-C., Iwahara, K., Watanabe, T, Honda, Y. and Kuwahara, M. (1996) Production and characterization of ligninolytic enzymes of Bjerkandera adusta grown on wood meal/wheat bran culture and production of these enzymes using a rotary-solid fermenter. Mycoscience 37, 417-425 https://doi.org/10.1007/BF02460998
  25. Ha, H.-C. and Lee, J.-S. (2002) froduction ot lignmoiytic enzymes from Pleurotus ostreatus grown on wood meal-wheat bran culture. J. Korean Soc. Agric. Chem. Biotechnol. 45, 124-127
  26. Kaal, E. E. J., Field, J. A. and Joyce, T. W. (1995) Increasing ligninolytic enzyme activities in several white-rot basidiomycetes by nitrogen-sufficient media. Bioresource Technol. 53, 133-139 https://doi.org/10.1016/0960-8524(95)00066-N
  27. Fu, S. Y, Yu, H.-S. and Buswell, J. A. (1997) Effect of nutrient nitrogen and manganese on manganese peroxidase and laccase production by Pleurotus sajor-caju. FEMS Microbiol. Lett. 147, 133-137 https://doi.org/10.1111/j.1574-6968.1997.tb10232.x
  28. Iwamoto, K., Ha, H.-C., Honda, Y, Watanabe, T. and Kuwahara, M. (1997) Isolation and characterization of manganese (II) Peroxidase (MnP) Produced by Pleurotus ostreatus. Wood Res. 84, 34-36
  29. Asada, Y, Watanabe, A., Irie, T., Nakarnura, Y. and Kuwahara, M. (1995) Structures of genomic and complementary DNAs coding for Pleurotus ostreatus manganese (II) Peroxidase. Biochem. Biophys. Acta 1251, 205-209
  30. Irie, T., Honda, Y, Ha, H.-C., Watanabe, T. and Kuwahara, M. (2000) Isolation of cDNA and genomic fragments encoding the major manganese peroxidase isoenzyme from the white rot basidiomycete Pleurotus ostreatus. J. Wood Sci. 46, 230-233 https://doi.org/10.1007/BF00776454
  31. Giardina, R, Palmieri, G., Fontanella, B., Rivieccio, V. and Sannia, G. (2000) Manganese peroxidase isoenzymes produced by Pleurotus ostreatus grown on wood sawdust. Arch. Biochem. Biophys. 376, 171-179 https://doi.org/10.1006/abbi.1999.1691
  32. Sarkar, S., Marti'nez, A. T. and Martinez, M. J. (1997) Biochemical and molecular characterization of a manganese peroxidase isoenzyme from Pleurotus ostreatus. Biochim. Biophys. Acta 1339, 23-30
  33. Wariishi, H., Valli, K. and Gold, M. H. (1992) Manganese (II) oxidation by manganese peroxidase from the basidiomycete Phanerochaete chrysosporium. J. Biol. Chem. 267, 23688-23695