Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification, Characterization, and cDNA Cloning of Xylanase from Fungus Trichoderma Strain SY

  • Min, Shin-Young (Department of Forest and Environmental Science & Bio/Molecular Informatics Center, Konkuk University) ;
  • Kim, Bong-Gyu (Department of Forest and Environmental Science & Bio/Molecular Informatics Center, Konkuk University) ;
  • Lee, Chan (Department of Food Science, Chungang University) ;
  • Hur, Hor-Gil (Department of Environmental Science and Engineering, Kwangju Institute of Science and Technology) ;
  • Ahn, Joong-Hoon (Department of Forest and Environmental Science & Bio/Molecular Informatics Center, Konkuk University)
  • Published : 2002.12.01
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Abstract

A xylanase-producing Trichoderma strain was isolated from soil. Xylanase from Trichoderma strain SY was purified 21-fold to an apparent homogeneity, with a $17.4\%$ yield. The optimum pH and temperature were determined to be 5.5 and $50^{\circ}C$, respectively, and its molecular weight was 21-kDa by SDS-PAGE. The corresponding gene, named xyl, was cloned by RT-PCR. DNA blot analysis of xyl showed that this gene is present as a single copy. The amino acid sequence of the Xyl protein showed similarity to those of other xylanases derived from various fungi. mRNA of xyl was highly expressed when this fungus was grown on cellulose or xylan as a sole carbon source, but undetectable when grown on sucrose. Extracts of Escherichia coli cells expressing xyl were found to have xylanase activity. It was confirmed that xyl from this isolate encodes xylanase.

Keywords

References

  1. Biochem. v.65 Molecular cloning and characterization of cel2 from the fungus Cochliobolus carbonum Ahn,J.H.;P.sposato;S.I.Kim;J.D.Walton
  2. Anal. Biochem. v.72 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Bradford,M. https://doi.org/10.1016/0003-2697(76)90527-3
  3. Fungal. Genet. Biol. v.25 Carbon catabolite repression of gene expression and conidiation in Neurospora crassa Ebbole,D.J. https://doi.org/10.1006/fgbi.1998.1088
  4. Plant Physiol. v.121 The enzymatic activity of fungal xylanase is not necessary for its elicitor activity Enkerli,J.;G.Felix;T.Boller https://doi.org/10.1104/pp.121.2.391
  5. Proc. Natl. Acad. Sci. v.85 Rapid production of full-length cDNAs from rare transcripts: Amplification using a single-specific oligonucleotide primer Frohman,M.A.;M.k.Dush;G.R.Martin https://doi.org/10.1073/pnas.85.23.8998
  6. Trichoderma and Gliocladium. v.I Morphology and identification of Trichoderma Games,W.;J.Bissett;Kubicek,C.P.;Harman,G.E.(eds.)
  7. Bioresource Tech v.58 Production of fungal xylanases Haltrich,D.;B.Nidetzky;K.D.Kulbe;W.Steiner;S.Zupancic https://doi.org/10.1016/S0960-8524(96)00094-6
  8. J. Microbiol. Biotechnol. v.11 Carbon catabolite repression (CCR) of expression of the xylanase A gene of Bacillus stearothermophilus No.236 Ha,G.S.;I.D.Choi;Y.J.Choi
  9. Gel electrophoresis of proteins: A Practical Approach Hames,B.D.;D.Rickwood
  10. Biochem. J. v.293 New familes in the classification of glycosyl hydrolases based on amino acid sequence similarities Henrissat,B.;A.Bairoch https://doi.org/10.1042/bj2930781
  11. Enzyme Microbiol. Technol. v.11 Cellulase and xylan degrading enzymes of Asperfillus terreus Hrmova,M.;P.Biely;M.Vrsanka https://doi.org/10.1016/0141-0229(89)90090-2
  12. J. Microbiol. Biotechnol. v.10 Conversion of xylose to ethanol by recombinant Saccharomyces cerevisiae containing genes for xylose reductase and xylitol dehydrogenase from Pichia stipitis Jin,Y.S.;T.H.Lee;Y.D.Choi;Y.W.Ryu;J.H.Seo
  13. J. Microbiol. Biotechnol. v.12 Cellulase-free thermostable alkaline xylanase from thermophilic and alkalophilic Bacillus sp. JB-99 Johnvesly,B.;S.Virupakshi;G.N.Patil;A.Ramalingam;G.R.Naik
  14. J. Microbiol. Biotechnol. v.10 Regulation of cycloinulooligosaccharide fructanotransferase synthesis in Bacillus macerans and Bacillus subtilis Kim,H.Y.;Y.J.Choi
  15. J. Microbiol. Biotechnol. v.10 Constitutive overexpression of the endoxylanase gene in Bacillus subtilis Kim,J.H.;J.H.Kim;S.C.Kim;S.W.Nam
  16. FEMS Microbiol. Rev. v.23 Molecular and biotechnological aspects of xylanases Kulkarni,N.;A.Shendye;M.Rao https://doi.org/10.1111/j.1574-6976.1999.tb00407.x
  17. J. Microbiol. Biotechnol. v.1 Pilot-scale production of cellulase using Trichoderma reesei rut C-30 in fed-batch mode Lee,S.M.;M.K.Yoon
  18. J. Microbiol. Biotechnol. v.10 Qualitative evaluation of ligninolytic enzymes in xylariaceous fungi Lee,Y.S.
  19. J. Microbiol. Biotechnol. v.11 Asparagine residue at position 71 is responsible for alkalitolerance of the xylanase from Bacillus pumilus A 30 Liu,X.M.;M.Qi;J.Q.Lin;Z.H.Wu;Y.B.Qu
  20. Biochem. J. v.239 Purification and properties of xylanases from Streptomyces lividans Morosoli,R.J.;J.L.Bertrand;F.Mondou;F.Shareck;D.Klupfel https://doi.org/10.1042/bj2390587
  21. Enzyme Microb. Technol. v.13 Enzymatic production of oligosaccharides from corncob xylan Pellerin,P.;M.Gosselin;J.P.Lepoutre;E.Samain;P.Deberie https://doi.org/10.1016/0141-0229(91)90074-K
  22. Cold Spring Harbor Laboratory Press (2? Ed.) Molecular cloning; A Laboratory Manual Sambrook J.;E.F.Fritsch;T.Maniatis
  23. J. Microbiol. Biotechnol v.11 Purification and characterization of a regulatory protein xy1R in the D-xylose operon from Escherichia coli Shin,J.H.;D.H.Roh;G.Y,Heo;G.J.Joo;I.K.Rhee
  24. Plant Cell v.12 The Cochliobolus carbonum SNF1 gene is required for cell wall-degrading enzyme expression and virulence on maize Tonukari,N.J.;J.S.Scott Craig;J.D.Walton https://doi.org/10.1105/tpc.12.2.237
  25. Physiol. Mol. Plant Path. v.39 A single β-1,3-glucanase secreted by the maize pathogen Cochliobolus carbonum acts by an exolytic mechanism Van Hoof,A.;J.Leykam;H.J.Schaeffer;J.D.Walton https://doi.org/10.1016/0885-5765(91)90034-F
  26. FEMS Microbiol. Rev. v.13 Xylanases in bleaching: From an idea to the industry Viikari,L.;A.Kantelinen;J.Sundquist;M.Linko https://doi.org/10.1111/j.1574-6976.1994.tb00053.x
  27. Plant Physiol v.104 Deconstructing the cell wall Walton,J.D. https://doi.org/10.1104/pp.104.4.1113