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Purification and characterization of dye degrading of veratryl alcohol oxidase from Pseudomonas aeruginosa strain BCH

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Abstract

In the present study we have purified the intracellular veratryl alcohol oxidase (VAO) enzyme from Pseudomonas aeruginosa strain BCH to evaluate its dye decolorizing potential. The enzyme was purified by ion exchange chromatography using DEAE cellulose followed by gel filtration chromatography using Biogel P-100. The molecular weight of the purified enzyme was estimated by polyacrylamide gel electrophoresis (PAGE) analysis. The VAO was purified up to 12 and 16.3-fold by ion exchange and gel filtration chromatography respectively. VAO was estimated to be about 85 kDa by SDS–PAGE. The optimum pH and temperature for purified VAO was 3 and 55°C respectively. The purified enzyme exerted its optimal activity with veratryl alcohol and also oxidized various other substrates, whereas diminished activity was noted in case of tryptophan and xylidine. The metal ions Mn++ and Hg++ were found to suppress the oxidase activity. The purified enzyme decolorized different dyes with variable decolorization rates and efficiencies. Decolorization mechanism of Remazol Black by purified enzyme was studies in detail using various analytical techniques like HPLC, GC–MS and FTIR. This study is useful for understanding the precise role of Pseudomonas aeruginosa strain BCH in the decolorization of textile dyes containing industrial wastewater.

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Acknowledgments

Swapnil S. Phugare one of the authors would like to thank to Jawaharlal Nehru Memorial Fund, New Delhi for awarding JNMF fellowship for doctoral studies.

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Authors and Affiliations

  1. Department of Biochemistry, Shivaji University, Kolhapur, 416 004, India

    Swapnil S. Phugare & Jyoti P. Jadhav

  2. Department of Microbiology, Shivaji University, Kolhapur, 416 004, India

    Shailesh R. Waghmare

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  1. Swapnil S. Phugare
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  2. Shailesh R. Waghmare
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  3. Jyoti P. Jadhav
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Correspondence to Jyoti P. Jadhav.

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Phugare, S.S., Waghmare, S.R. & Jadhav, J.P. Purification and characterization of dye degrading of veratryl alcohol oxidase from Pseudomonas aeruginosa strain BCH. World J Microbiol Biotechnol 27, 2415–2423 (2011). https://doi.org/10.1007/s11274-011-0714-6

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  • DOI: https://doi.org/10.1007/s11274-011-0714-6

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