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Redefining XynA from Penicillium funiculosum IMI 378536 as a GH7 cellobiohydrolase

  • Biocatalysis
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The secretome of Penicillium funiculosum contains two family GH7 enzymes, one of which (designated XynA) has been described as a xylanase. This is unusual because it is the only xylanase in family GH7, which is mainly composed of cellobiohydrolases and endoglucanases, and also because XynA is highly similar to the cellobiohydrolase I from Talaromyces emersonii and Trichoderma reesei (72 and 65 % identity, respectively). To probe this enigma, we investigated the biochemical properties of XynA, notably its activity on xylans and β-d-glucans. A highly pure sample of XynA was obtained and used to perform hydrolysis tests on polysaccharides. These revealed that XynA is 100-fold more active on β-1,4-glucan than on xylan. Likewise, XynA was active on both 4-nitrophenyl-β-d-lactopyranoside (pNP-β-d-Lac) and 4-nitrophenyl-β-d-cellobioside (pNP-cellobiose), which shows that XynA is principally an exo-acting type 1 cellobiohydrolase enzyme that displays 5.2-fold higher performance on pNP-cellobiose than on pNP-β-d-Lac. Finally, analyses performed using cellodextrins as substrate revealed that XynA mainly produced cellobiose (C2) from substrates containing three or more glucosyl subunits, and that C2 inhibits XynA at high concentrations (IC50 C2 = 17.7 μM). Overall, this study revealed that XynA displays typical cellobiohydrolase 1 activity and confirms that the description of this enzyme in public databases should be definitively amended. Moreover, the data provided here complete the information provided by a previous proteomics investigation and reveal that P. funiculosum secretes a complete set of cellulose-degrading enzymes.

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Acknowledgments

The authors would like to thank the Agence National de la Recherche et de la Technologie for financial support accorded to Hélène Texier. Dr. Monsarrat and his team at the IPBS (Institut de Pharmacologie et de Biologie Struturale, Toulouse, France) are thanked for the protein sequence and glycosylation analysis.

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Authors and Affiliations

  1. Université de Toulouse, INSA, UPS, INP; LISBP, 135 Avenue de Rangueil, 31077, Toulouse, France

    Hélène Texier, Claire Dumon & Michael J. O’Donohue

  2. INRA, UMR792, Ingénierie des Systèmes Biologiques et des Procédés, 31400, Toulouse Cedex 4, France

    Hélène Texier, Claire Dumon & Michael J. O’Donohue

  3. CNRS, UMR5504, 31400, Toulouse, France

    Hélène Texier, Claire Dumon & Michael J. O’Donohue

  4. Cinabio ADISSEO France SAS, Hall Gilbert Durand 3, 135 Avenue de Rangueil, 31077, Toulouse, France

    Hélène Texier, Virginie Neugnot-Roux & Marc Maestracci

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  1. Hélène Texier
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  2. Claire Dumon
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  3. Virginie Neugnot-Roux
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Correspondence to Michael J. O’Donohue.

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Texier, H., Dumon, C., Neugnot-Roux, V. et al. Redefining XynA from Penicillium funiculosum IMI 378536 as a GH7 cellobiohydrolase. J Ind Microbiol Biotechnol 39, 1569–1576 (2012). https://doi.org/10.1007/s10295-012-1166-1

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  • DOI: https://doi.org/10.1007/s10295-012-1166-1

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