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Purification and characterization of laccase produced by a white rot fungus Pleurotus sajor-caju under submerged culture condition and its potential in decolorization of azo dyes

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Abstract

An extracellular laccase was isolated and purified from Pleurotus sajor-caju grown in submerged culture in a bioreactor, and used to investigate its ability to decolorize three azo dyes. The extracellular laccase production was enhanced up to 2.5-fold in the medium amended with xylidine (1 mM). Purification was carried out using ammonium sulfate (70% w/v), DEAE-cellulose, and Sephadex G-100 column chromatography. The enzyme was purified up to 10.3-fold from the initial protein preparation with an overall yield of 53%. The purified laccase was monomeric with an apparent molecular mass of 61.0 kDa. The purified enzyme exerted its optimal activity with 2,2-azino–bis(3-ethylbenzo-thiazoline-6-sulfonate (ABTS) and oxidized various lignin-related phenols. The catalytic efficiencies k cat/K m determined for ABTS and syringaldazine were 9.2×105 and 8.7×105, respectively. The optimum pH and temperature for the purified enzyme was 5.0 and 40 °C, respectively. Sodium azide completely inhibited the laccase activity. The absorption spectrum revealed type 1 and type 3 copper signals. The purified enzyme decolorized azo dyes such as acid red 18, acid Black 1, and direct blue 71 up to 90, 87, and 72%, respectively. Decolorization ability of P. sajor-caju laccase suggests that this enzyme could be used for decolorization of industrial effluents.

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Authors and Affiliations

  1. Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai, Tamil Nadu 600 025, India

    Kumarasamy Murugesan, Manavalan Arulmani & P. Thangavelu Kalaichelvan

  2. School of Environmental Science and Engineering, Pohang University of Science and Technology, San 31, Hyojadong, Namgu, Pohang 790-784, South Korea

    Kumarasamy Murugesan, In-Hyun Nam, Young-Mo Kim & Yoon-Seok Chang

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  1. Kumarasamy Murugesan
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  2. Manavalan Arulmani
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  3. In-Hyun Nam
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  4. Young-Mo Kim
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  5. Yoon-Seok Chang
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  6. P. Thangavelu Kalaichelvan
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Correspondence to Yoon-Seok Chang or P. Thangavelu Kalaichelvan.

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Murugesan, K., Arulmani, M., Nam, IH. et al. Purification and characterization of laccase produced by a white rot fungus Pleurotus sajor-caju under submerged culture condition and its potential in decolorization of azo dyes. Appl Microbiol Biotechnol 72, 939–946 (2006). https://doi.org/10.1007/s00253-006-0403-9

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  • DOI: https://doi.org/10.1007/s00253-006-0403-9

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