Enzyme Activities and Cellulose Degradation of Domestic Softwoods in Shaking Culture of Fomitopsis palustris

국내산 침엽수 목분의 진탕배양에서 나타난 Fomitopsis palustris의 효소 활성 및 셀룰로오스 분해

  • Received : 2007.10.10
  • Accepted : 2007.10.24
  • Published : 2007.11.25

Abstract

Activities of the extracellular enzyme from Fomitopsis palustris, a brown-rot fungi, and by which crystallinity changes of cellulose in the various softwoods, such as Larix leptolepsis, Finns rigida, Finns koraiensis and Finns densiflora by liquid culture, were investigated. Activity of Cellobiohydrolase (CBH) from F. palustris was detected in the every test softwoods culture, showing activities of the Endoglucanase (EG), $\beta$-glucosidase (BGL) and $\beta$-1,4-xylosidase (BXL). It was shown high enzyme activities in the sapwood culture than heartwood of the same wood species, However, the enzyme activities in most of test wood cultures increased with longer incubation time, indicating a possibility of intermix sapwood and heartwood for degradation process by enzyme. Also it was shown that protein patterns of the extracellular enzyme from F. palustris in wood particle substrate of the several domestic softwoods were similar with each other wood species, which suggested the possibility of mixing all softwoods in saccharification by enzyme from F. palustris. Crystallinity reduction value of cellulose by F. palustris was 4.2~20.4% in 4 weeks cultivation, 12.9~28.9% in 8 weeks.

소나무, 낙엽송, 리기다소나무, 잣나무 등 국내산 침엽수 목분을 기질로 진탕배양했을 때 갈색부후균인 Fomitopsis palustris로부터 분비된 당 분해성 균체 외 효소는 Endoglucanase (EG), $\beta$-glucosidase (BGL) 및 $\beta$-1,4-xylosidase (BXL)와 함께 결정형 셀룰로오스를 분해하는 Cellobiohydrolase (CBH)도 활성을 갖은 것으로 나타났다. 4주간 배양에서 변재는 심재에 비해 큰 효소활성을 나타냈으나, 배양기간을 증가시킴으로써 효소활성이 커지는 것으로 밝혀져 효소에 의한 목질바이오매스 분해의 경우 심변재 혼합처리도 가능할 수 있음이 시사되었다. 그리고, 균체 의 단백질을 SDS-PAGE로 분석한 결과, 대부분의 수종에서 나타나는 효소의 단백질 패턴은 거의 유사한 것으로 나타나 효소를 이용한 목질계 바이오매스 분해의 경구 수종별 혼합처리도 가능함이 시사되었다. F. palustris에 의해 분해된 목분(60 mesh 통과) 셀룰로오스의 결정화도 감소율은 4주 배양에서 약 4.2~20.4%, 8주 배양에서 약 12.9~28.9% 수준으로 나타났다.

Keywords

References

  1. 김영숙, 김이경, 최두열, 윤정준, 김영균. 2006 침엽수 목질계 기질에서 나타난 Fomitopsis palustris의 효소 특성. 한국목재공학회 요지집. 50-51
  2. 金潤受. 2007. 褐色腐朽への再訪 一褐色腐朽の統さ れた概念は得まだられていない一. 木材保存. vol. 33-2
  3. 金潤受, 金圭赫, 金榮淑. 2004. 木材保存科學. 71-72. 全南大學校出版部
  4. 전문진, 권석태, 이철호, 임번삼. 2003. 현대의 생물공학과 생물산업. 아카데미서적
  5. 최지호, 한옥수, 김윤수. 1992. 갈색부후목재의 X선 회절 및 IR 분석. 목재공학. 20(3): 55 - 60
  6. Miieninen-Oinonen, A. 2004. Trichoderma reesei strains for production of celIulases for the textile industry. p22. VTT publications. 550
  7. Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of pro tein-dye binding, Anal. Biochem. 72: 248- 254 https://doi.org/10.1016/0003-2697(76)90527-3
  8. Flournoy, D. S., T. K. Kirk, and T. L. Highley. 1991. Wood decay by brown-rot fungi: Changes in pore structure and cell wall volume. Holzforschung. 45: 383-388 https://doi.org/10.1515/hfsg.1991.45.5.383
  9. Highly, T. L, R. Ibach, and T. K. Kirk 1988. Properties of cellulose degraded by the brown rot fungus Postia placenta. IRG/WP/Document No. 1350
  10. Ishihara, M. and K. Shimizu. 1984a. Chemical characteristics of brown rot Tyromyces palustris. Bull. Jap, For. & For. Prod. Res. Inst. No. 330: 141-152
  11. Ishihara, M. and K. Shimizu. 1984b. Purification and properties of two extracellular endo-cellulases from the brown rotting fungus Tyromyces palustris. Mokkuzai Gakkaishi. 30: 79-87
  12. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227: 680-685 https://doi.org/10.1038/227680a0
  13. Reczey, K. 1996. Cellulase production by T.reesei
  14. Keller, F. A., J. E. Hamilton, and Q. A. Nguyen. 2003. Microbial pretreatment of biomass, Applied Biochemistry and Biotechnology. vol. 105-108: 27-85
  15. McMillan, J. D. 2003. Enzyme sugar-ethanol platform and advanced pretreatment interim project reviews. http://www.nrel.gov/bioenergy
  16. Segal, L., J. J. Creely, A. E. Jr. Martin, and C. M. Conrad. 1959. An empirical method for estimating the degree of crystallinity of native cellulose using the X-ray diffractometer. Text. Res. J. 786-794
  17. Somogyi, M. 1959. Exacerbation of diabetes by excess of insulin action. Am. J. Med. 26: 169 https://doi.org/10.1016/0002-9343(59)90307-9
  18. Kim, Y.-S., J.-J. Yoon, N. Khan, Y.-K. Kim, C.-J. Cha, and Y.-K. Kim. 2006. Hydrolysis of crystalline cellulose by cellulase system from the brown-rot fungus Fomitopsis palustris. 2006 Proceedings of the Korean Society of Wood Science and Technology Annual meeting. 48-49
  19. Yoon, J.-J. and Y.-K. Kim. 2005. Degradation of Crystalline Cellulose by the Brown-rot Basidiomycete F. palustris. Journal of Microbiology. December 2005. pp. 487-492
  20. Yoon, J.-J., C.-J. Cha, Y.-S. Kim, and D.-W. Son. 2007. The brown-rot basidiomycete F. palustris has the Endo-glucanases capable of degrading microcrystalline cellulose. J. Microbial Biotechnol. 17(5): 800-805
  21. Zabel, R. A. 1992. Wood microbiology. pp. 204-208. Academic press