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Production of a chimeric enzyme tool associating the Trichoderma reesei swollenin with the Aspergillus niger feruloyl esterase A for release of ferulic acid

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Abstract

The main goals of this work were to produce the fusion protein of the Trichoderma reesei swollenin I (SWOI) and Aspergillus niger feruloyl esterase A (FAEA) and to study the effect of the physical association of the fusion partners on the efficiency of the enzyme. The fusion protein was produced up to 25 mg l−1 in the T. reesei strains Rut-C30 and CL847. In parallel, FAEA alone was produced for use as a control protein in application tests. Recombinant FAEA and SWOI–FAEA were purified to homogeneity and characterized. The biochemical and kinetic characteristics of the two recombinant proteins were found to be similar to those of native FAEA, except for the temperature stability and specific activity of the SWOI–FAEA. Finally, the SWOI–FAEA protein was tested for release of ferulic acid from wheat bran. A period of 24 h of enzymatic hydrolysis with the SWOI–FAEA improved the efficiency of ferulic acid release by 50% compared with the results obtained using the free FAEA and SWOI. Ferulic acid is used as an antioxidant and flavor precursor in the food and pharmaceutical industries. This is the first report of a potential application of the SWOI protein fused with an enzyme of industrial interest.

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Acknowledgements

This research was supported by the sixth European Framework Program for Research and Technological Development (FP6), Integrated Project No. 019882; NILE project: New Improvements for Lignocellulosic Ethanol; and the French Programme National de Recherche sur les Bioénergies (PNRB), SPECABBE project, Stratégie de Prétraitements Physiques, Enzymatiques, et Chimiques Appliquées à la Biomasse–Bio-Ethanol.

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Authors and Affiliations

  1. UMR 1163 INRA/Universités de Provence et de la Méditerranée de Biotechnologie des Champignons Filamenteux, IFR-IBAIM, 163 avenue de Luminy, Case Postale 925, 13288, Marseille, Cedex 09, France

    Anthony Levasseur, David Navarro, Marcel Asther & Eric Record

  2. VTT Technical Research Centre of Finland, P.O. Box 1500, Tietotie 2, 02044 VTT, Espoo, Finland

    Markku Saloheimo, Martina Andberg & Tiina Nakari-Setälä

  3. IFP/Département de Biotechnologie et Chimie de la Biomasse, 1-4 avenue de Bois-Préau, 92852, Rueil-Malmaison, cedex, France

    Frédéric Monot

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  1. Anthony Levasseur
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  2. Markku Saloheimo
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  3. David Navarro
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  4. Martina Andberg
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  5. Frédéric Monot
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  6. Tiina Nakari-Setälä
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  7. Marcel Asther
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  8. Eric Record
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Correspondence to Eric Record.

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Levasseur, A., Saloheimo, M., Navarro, D. et al. Production of a chimeric enzyme tool associating the Trichoderma reesei swollenin with the Aspergillus niger feruloyl esterase A for release of ferulic acid. Appl Microbiol Biotechnol 73, 872–880 (2006). https://doi.org/10.1007/s00253-006-0546-8

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  • DOI: https://doi.org/10.1007/s00253-006-0546-8

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