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The Two Major Xylanases from Trichoderma Reesei: Characterization of Both Enzymes and Genes

Bio/Technology volume 10, pages 1461–1465 (1992)Cite this article

Abstract

As a first step to exploit the potential of Trichoderma reesei to produce hemicellulases, we have purified two endo-β-1,4-xylanases (1,4-β-D-xylan xylanohydrolase, EC 3.2.1.8) and cloned their genes. The enzymes were isolated from culture filtrates of T. reesei C 30 grown on xylan as a carbon source, using two steps of cation exchange chromatography. They exhibited molecular weights of 19 (XYN I) and 21 (XYN II) kD, and isoelectric points of 5.2 and 9.0, respectively. These enzymes differed in their pH optimum for activity and affinity for xylan, and accounted for more than 90% of the total xylanolytic activity of the fungus. The purified enzymes were subjected to N-terminal sequence analysis, and after cleavage with trypsin and endoproteinase Glu-C the resulting peptides were sequenced. Oligonucleotides based on these sequences were used to clone gene fragments via PCR, and these were used as probes to isolate full-length copies of xyn1 and xyn2 from a lambda gene bank of T. reesei. The products of xyn1 and xyn2 share considerable homology, but the enzyme encoded by xyn2 appears to more closely resemble several other bacterial and fungal xylanases than does that of xyn1.

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Author notes

  1. Ramon Gonzalez & Christian P. Kubicek

    Present address: CSIC, Instituto de Agroquimica y Tecnologia de Alimentos, Valencia, Spain

  2. Christian P. Kubicek: Corresponding author.

Authors and Affiliations

  1. Abteilung für Mikrobielle Biochemie, Institut für Biochemische Technologie und Mikrobiologie, TU Wien, Getreidemarkt 9, A-1060, Wien, Austria

    Anneli Törrönen, Robert L. Mach & Robert Messner

  2. Institute of Biotechnology, University of Helsinki, 00380, Helsinki, Finland

    Nisse Kalkkinen

  3. Cultor Ltd. Technology Center, Kantvik, SF-2460, Finland

    Anu Harkki

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Törrönen, A., Mach, R., Messner, R. et al. The Two Major Xylanases from Trichoderma Reesei: Characterization of Both Enzymes and Genes. Nat Biotechnol 10, 1461–1465 (1992). https://doi.org/10.1038/nbt1192-1461

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  • DOI: https://doi.org/10.1038/nbt1192-1461

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