The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
권호 표지

Production of Alkaline Carboxymethyl Cellulase and Xylanase by Batch and Fed-batch Cultures of Alkalophilic Cephalosporium sp. RYM-202

호알카리성 Cephalosporium sp. RYM-202의 회분 및 유가배양에 의한 Alkaline Carboxymethyl Cellulase와 Xylanase의 생산

  • Kang, Myoung-Kyu (Department of Environmental Technology, Tonghae College) ;
  • Kim, Do-Young (Department of Microbiology, Chungnam National University) ;
  • Rhee, Young-Ha (Department of Microbiology, Chungnam National University)
  • 강명규 (동해전문대학 환경공업과) ;
  • 김도영 (충남대학교 자연과학대학 미생물학과) ;
  • 이영하 (충남대학교 자연과학대학 미생물학과)
  • Published : 1997.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Production of alkaline carboxymethyl cellulase (CMCase) and xylanase by batch and fed-batch cultures of alkalophilic Cephalosporium sp. RYM-202 was investigated. Of carbon sources tested, wheat bran gave the highest production of those enzymes. The high levels of CMCase on carboxymethyl cellulose and xylanase on birchwood xylan suggest that the biosynthesis of CMCase and xylanase in Cephalosporium sp. RYM-202 is regulated separately at the level of enzyme induction. The temperature and pH for maximal production of those enzymes was $20^{\circ}C$ and 9.0, respectively. High concentration of wheat bran in batch fermentation resulted in the lower and delayed production of the enzymes by catabolite repression. In fed-batch fermentation with controlled feeding of 5% final wheat bran concentration, the highest activities of CMCase and xylanase were 0.39 and 9.2 units/ml, respectively, and 1.22 and 1.36 times higher respectively than those in batch fermentation on 5% wheat bran.

회분배양과 유가배양을 이용하여 호알칼리성 Cephalosporium sp. RYM-202로부터 alkaline carboxymethyl cellulase (CMCase)와 Xylanase의 생산을 위한 배양조건에 대하여 조사하였다. 조사한 탄소기질 중에서 밀기울이 두 효소의 생산에 가장 효율적이었다. 또한 CMCase는 carboxymethyl cellulose (CMC)를 탄소기질로 첨가한 배양액에서, 반면에 xylanase의 경우에는 xylan을 기질로 하였을 때 높은 생산량을 나타냄으로서 유도기질 특이성을 보였는 바, 이 결과는 Cephalosporium sp. RYM-202에서의 CMCaee와 xylanase의 생합성이 효소 유도 수준에서 독립적으로 조절됨을 시사해 준다. 조사된 질소원 중에서는 무기질소원인 $NaNO_3$가 효소생산에 효과적이었으며, 이들 효소의 최대 생산을 위한 배양온도와 pH는 각각 $20^{\circ}C$와 9.0인 것으로 나타났다. 한편, 발효조에서의 회분배양을 통한 효소생산의 경우, 밀기울의 농도를 5%까지 증가시킴에 따라 효소생산량은 증가되었으나 catabolite repression에 의해 효소생산의 지연과 생산력의 감소를 초래하였다. 이러한 문제점은 탄소원의 간헐적 공급에 의한 유가배양을 통해서 어느 정도 해결될 수 있는 것으로 나타났으며, 밀기울의 최종농도가 5% 되게 공급된 유가배양 시 CMCase와 xylanase의 최대 효소생산량은 각각 0.39 및 9.2 units/ml 이었으며, 이는 같은 농도의 밀기울을 함유하는 회분배양 시 획득된 효소활성에 비해 각각 1.22배와 1.36배 증가된 것이다.

Keywords

References

  1. Appl. Microbiol. Biotechnol v.44 Optimization of extracellular xylanase production by Dictyoglomus sp. Bl in continuous culture Adamsen, A.K.;Lindhagen, J.;Ahring, B.K.
  2. Can. J. Microbiol. v.31 Production, purification, and properties of thermostable xylanase from Clostridium stercorarium Berenger, J.F.;Frixon, C.;Bigliardi, J.;Creuzet, N.
  3. Enzyme Microb. Technol v.9 Development of a medium for high cellulase, xylanase and ${\beta}$-glucosidase production by a mutant strain (NTG III/6) of the cellulolytic fungus Penicillium pinophilum Brown, J.A.;Collin, S.A.;Wood, T.M.
  4. Biotechnol. Bioeng. v.27 Novel process for the production of cellulolytic enzymes Cauchon, N.;LeDuy, A.
  5. Appl. Microbiol. Biotechnol v.43 Optimization of cellulase production by Penicillium occitanis Chaabouni, S.E.;Belguith, H.;Hassairi, I.;M'Rad, K.;Ellouz, R.
  6. Can. J. Microbiol v.38 Purification and properties of extracellular endoxylanase from alkalophilic thermophilic Bacillus sp Dey, D.;Hinge, J.;Shendye, A.;Rao, M.
  7. J. Gen. Microbiol v.139 Bacterial cellulases and xylanases Gilbert, H.J.;Hazlewood, G.P.
  8. J. Gen. Microbiol. v.135 Regulation of the production of hemicellulolytic and cellulolytic enzymes by a Streptomyces sp. growing on lignocellulose Godden, B.;Legon, T.;Helvenstein, P.;Penninckx, M.
  9. Appl. Microbiol. Biotechnol. v.36 Production of cellulase and xylanase by a wild strain of Trichoderma viride Gomes, I.;Gomes, J.;Steiner, W.;Esterbauer, H.
  10. Appl. Microbiol. Biotechnol v.39 Production of a high level of cellulase-free xylanase by the thermophilic fungus Thermomyces lanuginosus in laboratory and pilot scales using lignocellulosic materials Gomes, J.;Purkarthofer, H.;Hayn, M.;Kapplmuller, J.;Sinner, M.;Steiner, W.
  11. Biotechnol. Lett. v.4 Enhancement of cellulase production by Trichoderma viride using carbon/nitrogen double-fed-batch Gottvaldova, M.;Kucera, J.;Podrazky, V.
  12. TIBTECH. v.10 A perspective on the biotechnological potential of extreamophiles Herbert, R.A.
  13. Enzyme. Microbiol. Technol v.6 Enhanced celluJase production in fed-batch culture of Trichoderma reesei C-30 Hendy, N.A.;Wilke, C.R.;Blanch, H.W.
  14. Appl. Microbiol. Biotechnol v.43 Cellulasefree xylanase by thermophilic fungi: a comparison of xylanase production by two Thermomyces lanuginosus strains Hoq, M.M.;Deckwer, W.D.
  15. Enzyme Microb. Technol. v.11 Cellulose and xylan-degrading enzymes of Aspergillus terreus and Aspergillus niger Hrmova, M.;Biely, P.;Vrsanska, M.
  16. J. Gen. Micribiol v.137 Induction of cellulose- and xylan-degrading enzyme systems in Aspergillus terreus by homo- and heterodisaccharides composed of glucose and xylose Hrmova, M.;Petrakova, E.;Biely, P.
  17. Agric. Bioi. Chem. v.53 Alkaline cellulase for laundry detergents: production by Bacillus sp. KSM？635 and enzymatic properties Ito, S.;Shikata, S.;Ozaki, K.;Kawai, S.;Okamoto, K.;Inoue, S.;Takei, A.;Ohta, Y.I.;Satoh, T.
  18. Kor. J. Mycol. v.21 Partial purification and some properties of carboxymethyl cellulases from Cephalosporium sp. RYM-202 Kang, M.K.;Park, H.M.;Rhee, Y.H.;Kim, Y.S.;Kim, Y.K.
  19. J. Microbiol v.33 Purification and characterization of a xylanase from alkalophilic Cephalosporium sp. RYM-202 Kang, M.K.;Kwon, T.I.;Yang, Y.K.;Rhee, Y.H.
  20. Appl. Environ. Microbiol v.62 Purification and characterization of Two xylanases from alkalophilic Cephalosporium sp. strain RYM-202 Kang, M.K.;Maeng, P.J.;Rhee, Y.H.
  21. Biotechnol. Lett. v.17 Carboxymethyl cellulases active and stable at alkaline pH from alkalophilic Cephalosporium sp. RYM-202 Kang, M.K.;Rhee, Y.H.
  22. Crit. Rev. Biotechnol v.13 Lignocellulose biotechnology:current and future prospects Kuhad, R.C.;Singh, A.
  23. Kor. J. Mycol. v.17 Synergistic effect of substrates on the biosysthesis of cellulase and xylanase complexes from Aspergillus nidulans Lee, J.A.;Maeng, J.S.;Maeng, P.J.;Rhee, Y.H.
  24. Appl. Microbiol. Biotechnol v.43 Effect of agitation on growth and enzyme production of Trichoderma reesei in batch fermentation Lejeune, R.;Baron, G.V.
  25. Anal. Chem v.31 Use of dinitrosalicylic acid reagent for determination of reducing sugar Miller, G.L.
  26. Appl. Environ. Microbiol v.59 Purification and some properties of an alkaline xylanase from alkalophilic Bacillus sp. strain 41M-1 Nakamura, S.;Wakabayashi, K.;Nakai, R.;Aono, R.;Horikoshi, K.
  27. FEMS Microbiol. Lett. v.115 Xylanase production in Aspergillus nidulans: Induction and carbon catabolite repression Pinaga, P.;Fernandez-Espinar, M.T.;Valles, S.;Ramon, D.
  28. Appl. Microbiol. Biotechnol v.37 Production of xylanolytic enzymes by an alkalitolerant Bacillus circulans strain Ratto, M.;Poutanen, K.;Viikari, L.
  29. Kor. J. Mycol. v.18 Influence of substrates on the isozyme patterns of cellulase and xylanase complexes in Aspergillus niger Rho, J.R.;Rhee, Y.H.;Chung, J.H.
  30. Kor. J. Microbiol v.31 Purification and characterization of xylanases from alkalophilic Streptomyces sp. S-510 Rhyum, S.B.;Kang, M.K.;Maeng, P.J.;Park, H.M.;Rhee, Y.H.
  31. Appl. Environ. Microbiol v.56 Interrelationship of xylanase induction and cellulase induction of Trichoderma longibrachiatum Royer, J.C.;Nakas, J.P.
  32. Biotechnol. Bioeng. v.37 The interaction of xylanases with commercial pulp Senior, D.J.;Mayers, P.R.;Saddler, J.N.
  33. Biotechnol. Bioeng v.25 Development of a culture medium for large scale production of cellulolytic enzymes by Trichoderma reesei Warzywoda, V.F.;Pourquie, J.
  34. Crit. Rev. Biotechnol v.12 Trichoderma xylanases, their properties and application Wong, K.K.Y.;Saddler, J.N.
  35. Fed-batch techniques in microbial processes;Advances in Biochemical Engineering/Biotechnology. Vol. 30 Yamane, T.;Shimizu, S.;Fiechter, A.(ed.)