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Purification and biochemical characterization of a new alkali-stable laccase from Trametes sp. isolated in Tunisia: role of the enzyme in olive mill waste water treatment

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Abstract

A white-rot basidiomycete, isolated from decayed acacia wood (from Northwest of Tunisia) and identified as Trametes sp, was selected in a broad plate screening because of its ability to decolorize and dephenolize olive oil mill wastewater (OMW) efficiently. The major laccase was purified and characterized as a monomeric protein with apparent molecular mass of 61 kDa (SDS-PAGE). It exhibits high enzyme activity over broad pH and temperature ranges with optimum activity at pH 4.0 and a temperature of 60 °C. The purified laccase is stable at alkaline pH values. The enzyme retained 50 % of its activity after 90 min of incubation at 55 °C. Using ABTS, this laccase presented K m and V max values of 0.05 mM and 212.73 μmoL min−1 mg−1, respectively. It has shown a degrading activity towards a variety of phenolic compounds. The purified laccase was partially inhibited by Fe2+, Zn2+, Cd2+ and Mn2+, while Cu2+ acted as inducer. EDTA (10 mM) and NaN3 (10 mM) were found to completely inhibit its activity. 73 % OMW was dephenolized after 315 min incubation at 30 °C with 2 U mL−1 of laccase and 2 mM HBT.

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Acknowledgments

This work has been carried out with funding from the Spanish MINECO (PRI-PIBAR-2011-1402) and CSIC (20120E106). Part of this work and fellowship was funded by the ministry of higher education and scientific research of Tunisia.

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Authors and Affiliations

  1. Laboratory of Enzyme Engineering and Microbiology, Ecole Nationale d’Ingénieurs de Sfax, University of Sfax, Route de Soukra Km 4,5, BP 1173, 3038, Sfax, Tunisia

    Dalel Daâssi, Héla Zouari-Mechichi, Moncef Nasri & Tahar Mechichi

  2. Centro de Investigaciones Biológicas (CIB-CSIC), Ramiro de Maeztu 9, 28040, Madrid, Spain

    Alicia Prieto & María Jesús Martínez

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  1. Dalel Daâssi
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  2. Héla Zouari-Mechichi
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  3. Alicia Prieto
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  4. María Jesús Martínez
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  5. Moncef Nasri
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  6. Tahar Mechichi
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Correspondence to Tahar Mechichi.

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Daâssi, D., Zouari-Mechichi, H., Prieto, A. et al. Purification and biochemical characterization of a new alkali-stable laccase from Trametes sp. isolated in Tunisia: role of the enzyme in olive mill waste water treatment. World J Microbiol Biotechnol 29, 2145–2155 (2013). https://doi.org/10.1007/s11274-013-1380-7

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