The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
권호 표지

Isolation and Characterization of Regulatory Mutant for Cellulase Production from Trichoderma reesei QM 9414

Trichoderma reesei QM 9414의 섬유소 분해 호소 생산을 위한 조절변이주의 분리 및 특성에 관한 연구

  • Choi, Kun-Ho (Department of Biological Engineering, Inha University) ;
  • Koo, Youn-Mo (Department of Biological Engineering, Inha University) ;
  • So, Jae-Seong (Department of Biological Engineering, Inha University)
  • Published : 1998.03.30
Download PDF
⟨ Previous Next ⟩

Abstract

Two regulatory mutants of Trichoderma reesei QM 9414 were isolated by treatment with N-methyl-N'-nitro-N-nitrosoguanidine, and the effects of various inducers on the carboxymethyIcellulose (CMC) and filter paper (FP) production were investigated. Induction of CMCase and FPase production of mutants was shown higher level than wild type strain in 1% lactose minimal broth. When induced by glucose, wild type showed glucose-repression for CMCase and FPase production and mutants showed glucose-derepression. Mutant 1 showed 8.38 fold higher CMCase activity and 5.68 fold higher FPase activity than wild type stain. Mutant 2 showed about 8.42 fold higher CMCase activity and 5.41 fold higher FPase activity than wild type strain. Enzyme activities from the mutants and wild type had the same optimum pH of 4.8 and optimum temperature of $60^{\circ}C$.

Trichoderma reesei QM 9414를 N-methyl-N'-nitro-N-nitrosoguanidine으로 처리하여 얻은 돌연변이주들을 여러 탄소원에서 배양했을 때 carboxymethylcellulose(CMC), fiter paper 등의 기질에 대한 활성측정결과 다음과 같은 결과를 얻었다. 모균주가 catabolite repression을 받는 반면에 돌연변이주들의 cellulase 활성은 glucose를 탄소원으로 사용한 배지에서 모균주에 비해 CMCase 8.4배, FPase가 $5.4{\sim}5.7$배 증가함으로써 glucose-derepression성질을 갖는 돌연변이주들을 얻었으며 glucose를 탄소원으로 사용하고 lactose로 효소의 생산을 유도시켜본 결과 모균주에 비해 효소의 생산에 안정성을 갖는 것으로 나타났으며 돌연변이주 1이 상대적으로 더 우수한 돌연변이주로 나타났다. 돌연변이주들에 의해 생산된 효소는 모균주와 마찬가지로 pH 4.8, $60^{\circ}C$에서 최적활성을 나타내었다.

Keywords

References

  1. 섬유소 분해효소 생산을 위한 조절 변이주의 개발 및 특성에 관한 연구 이철한
  2. 한국균학회지 v.24 no.3 고온성 변이균주 Talaromyces luteus 2004의 분리와 Carboxymethylcellulase의 생성 조절 및 효소의 특성 홍미경;한효영;정영희;민경희
  3. Appl. Microbiol. Biotechnol. v.46 Saccharomyces cerevisiae mutants selected for increased production of Trichoderma reesei cellulases Aho, S.;Arffman, A.;Korhola, M.
  4. Bio/Technology v.5 Cloning, characterization, and expression in Saccharomyces cerevisiae of endoglucanase I from Trichoderma reesei Arsdell, J.N.V.;Kwok, S.;Schweickart, V.L.;Ladner, M.;Gelfland, D.H.;Innis, M.A.
  5. Analy. Biochem. v.131 Detection of cellulase Activity in Polyacrylamine Gels Using Congo Red-Stained Agar Replicas Beguin, P.
  6. Anal. Biochem. v.72 A rapid and sensitive method for quantification of microgram quantities of protein utilizing the principle of protein-dye binding Bradford, M.M.
  7. Appl. Microbiol. Biotechnol. v.40 Trichoderma reesei cellulases in the bleaching of kraft pulp Buchert, J.;Ranua, M.;Siika-aho, M.;Pere, J.;Viikari, L.
  8. Enzyme. Microb. Technol. v.10 Genetic improvement of Trichoderma reesei for large scale cellulase production Durand, H.;Clanet, M.;Tiraby, G.
  9. Proc. Natl. Acad. Sci. USA v.86 Mechanism by which cellulose triggers cellobiohydrolase I gene expression in Trichoderma reesei El-Gogary, S.;Leite, A.;Crivellaro, O.;Eveleigh, D.E.;El-Dorry, H.
  10. Appl. Environ. Microbiol. v.47 no.1 Cellulase Biosysthesis in a Catabolite Repression-Resistant Mutant of Thermomonospora curvata Fennington, G.;Neubauer, D.;Stutzenberger, F.
  11. Microbiol. Biotechnol. Cellulase-poor xylanases produced by Trichoderma reesei RUT-C30 on hemicellulose substrates Gamerith, G.;Groicher, R.;Zeilinger, S.;Herzog, P.;Kubicek, C.
  12. Biotechnology Letters v.13 no.7 Production of Glucoamylase by 2-deoxy-D-glucose Resistant Mutant of Aspergillus Terreus 4 Ghosh, A.;Chatterjee, B.;Das, A.
  13. Appl. Environ. Microbiol. v.63 no.4 Regulation of Cellulase Gene Expression in the Filamentous Fungus Trichoderma reesei Ilmen, M.;Saloheimo, A.;Onnela, M.;Penttila, M.
  14. Appl. Environ. Microbiol. v.62 no.8 The Cellulases Endoglucanase I and Cellobiohydrolase II of Trichoderma reesei Act Synergistically To Solubilize Native Cotton Cellulose but Not To Decrease Its Molecular Size Kleman-leyer, K.;Siika-aho, M.;Teeri, T.;Kirk, T.
  15. Appl. Environ. Microbiol. v.62 no.8 Glucose-Induced Secretion of Trichoderma reesei Xylanases Kurzatkowski, W.;Torronen, A.;Filipek, J.;Mach, R.;Herzog, P.;Sowka, S.;Kubicek, C.P.
  16. Appl. Microbiol. Biotechnol. v.44 Cellulase induction by lactose in Trichoderma reesei PC-3-7 Morikawa, Y.;Ohashi, T.;Mantani, O.;Okada, H.
  17. Appl. Environ. Microbiol. v.61 no.10 Production of Trichoderma reesei Cellulases on Glucose-Containing Media Nakari-Setala, T.;Penttila, M.
  18. Biotechnology v.42 The biochemical nature of the cell envelope of a high cellulase-secreting mutant differs from that of the Trichoderma reesei wild type Nevalainen, H.;Lavygina, I.;Neethling, D.;Packer, N.
  19. Enzyme. Microbiol. Technol. v.9 Intraspecific hybridization of Trichoderma reesei QM 9414 by protoplast fusion using colour mutants Ogawa, K.;Brown, J.A.;Wood, T.M.
  20. Biotechnol Bioeng. v.32 Viscosity-enhancing bleaching of hardwood kraft pulp with xylanase from a cloned gene Paice, M.G.;Bernier, R.;Jurasek, L.
  21. Gene v.45 Homology between cellulase genes of Trichoderma reesei: complete nucleotide sequences of the endoglucanase I gene Penttila, M.;Lehtovaara, P.;Nevalainen, H.;Bhikhabhai, R.;Knowles, J.
  22. Appl. Microbiol. Biotechnol. v.42 Isolation of a high-specifie-growth-rate mutant of Cellulomonas flavigena on sugar cane bagasse Ponce-Noyola, T.;Torre, M.
  23. Science v.249 Three-Dimensional Structure of Cellobiohydrolase II from Trichoderma reesei Rouvinen, J.;Bergfors, T.;Teeri, T.;Knowles, J.K.C.;Jones, T.A.
  24. Appl. Microbiol. Biotechnol. v.41 Development of a transformation system for Trichoderma longibrachiatum and its use for constructing multicopy transformants for the egl1 gene Sanchez-Torres, P.;Gonzalez, R.;Perez-Gonzalez, J.A.;Gonzalez-Candelas, L.;Ramon, D.
  25. Bio / Technology v.1 Molecular cloning of exocellobiohydrolase I derived from Trichoderma reesei L 27 Shoemaker, S.;Schweickart, V.;Ladner, M.;Gelfand, D.;Kovak, S.;Myambo, K.;Innis, M.
  26. J. Bacteriol. v.123 no.2 Derepressed Synthesis of Cellulase by Cellulomonas Stewart, B.;Leatherwood, J.M.
  27. Appl. Environ. Microbiol. v.43 no.4 Use of Congo Red-Polysaccharide Interactions in Enumeration and Characterization of cellulolytic Bacteria from the Bovine Rumen Teather, R.;Wood, P.
  28. Biotechnolgy and Bioengineering v.24 Improvement of T. reesei Strain through Mutation and Selective Screening Techniques Wiley, J.
  29. Microbial Degradation of Natural Products Winkelmann, G.
  30. Bio / Technology v.6 Wood Hydrolysis by Cellulomonas Fimi Endoglucanase and Exoglucanase Coexpressed as Secreted Enzymes in Saccharomyces Cerevisiae Wong, W.K.R.;Curry, C.;Parekh, R.S.;Parekh, S.R.;Wayman, M.;Davies, R.W.;Kilburn, D.G.;Skipper, N.
  31. Methods Enzymol. v.160 Use of complex formation between Congo Red and polysaccharides in detection and assay of polysaccharide hydrolases Wood, P.J.;Erfle, J.D.;Teather, R.M.