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Purification and Properties of a Thermostable Xylanase GH 11 from Penicillium occitanis Pol6

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Abstract

An extracellular, endo-β-1,4-xylanase was purified to homogeneity from the culture filtrate of the filamentous fungus Penicillium occitanis Pol6, grown on oat spelt xylan. The purified enzyme (PoXyn2) showed a single band on SDS–PAGE with an apparent molecular weight of 30 kDa. The xylanase activity was optimal at pH 3.0 and 65 °C. The specific activity measured for oat spelt xylan was 2,368 U mg−1. The apparent K m and V max values were 8.33 mg ml−1 and 58.82 μmol min−1 ml−1, respectively, as measured on oat spelt xylan. Thin-layer chromatography experiments revealed that purified PoXyn2 degrades xylan in an endo-fashion releasing xylobiose as main end product. The genomic DNA and cDNA encoding this protein were cloned and sequenced. This PoXyn2 presents an open reading frame of 962 bp, not interrupted by any introns and encoding for a mature protein of 320 amino acids and 29.88 kDa.

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Acknowledgments

We express our gratitude to Prof. Gérard Tiraby and Dr. Henry Durand (Cayla Company, France) for kindly supplying the Penicillium occitanis (Pol6) strain used in this work. We are also grateful to Prof. Radhouane Ellouz for his stimulating and beneficial discussion. We thank Prof. Hafedh Mejdoub (FSS) for the sequencing of the NH2-terminus. We also thank Dr. Mohamed Eljawa (CBS) for the DNA sequencing. This work was supported financially by the “Ministère de l’Enseignement Supérieur, de la Recherche Scientifique et de la Technologie-Tunisia” through a grant to “Unité Enzyme et Bioconversion–ENIS.”

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  1. Unité Enzymes et Bioconversions, Ecole Nationale d’Ingénieurs de Sfax, University of Sfax, BP 1173, 3038, Sfax, Tunisia

    Dorra Driss, Fatma Bhiri, Mariem Siela, Raoudha Ghorbel & Semia Ellouz Chaabouni

  2. Unité de service Bioreacteur couplé à un Ultrafiltre, Ecole Nationale d’Ingénieurs de Sfax, University of Sfax, BP 1173, 3038, Sfax, Tunisia

    Fatma Bhiri, Raoudha Ghorbel & Semia Ellouz Chaabouni

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Correspondence to Dorra Driss.

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Driss, D., Bhiri, F., Siela, M. et al. Purification and Properties of a Thermostable Xylanase GH 11 from Penicillium occitanis Pol6. Appl Biochem Biotechnol 168, 851–863 (2012). https://doi.org/10.1007/s12010-012-9824-3

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