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The Determination of Assay for Laccase of Bacillus subtilis WPI with Two Classes of Chemical Compounds as Substrates

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Abstract

Ligninolytic enzyme complexes are involved in lignin degradation. Among them laccases are outstanding because they use molecular oxygen as a co-substrate instead of hydrogen peroxide as used by peroxidases. Bacterial laccase of Bacillus genus was first reported in Claus and Filip (Microbiol Res 152:209–216, 1997), since then more bacterial laccases have been found. In this research, laccase-producing bacteria were screened from pulp and paper industry wastewater, bagass and sugarcane rhizosphere. Nutrient agar medium containing 0.5 mM of guaiacol was used. It was observed that the laccase-producing strains developed brown colour from which 16 strains of Bacillus were identified. One of the isolated strains was identified as Bacillus subtilis WPI based on the results of biochemical tests and 16S rDNA sequence analysis. This strain showed laccase-like activity towards the oxidizing substrates ABTS and guaiacol. In this study guaiacol was used as the substrate of laccase activity assay. For determination of laccase activity of this isolate guaiacol was used as a substrate of assay for the first time in this study. SDS-PAGE and Native-PAGE confirmed the presence of laccase.

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Acknowledgments

This research was supported by Biotechnology and Biological Research Center of Shahid Chamran University of Ahvaz (Iran).

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Authors and Affiliations

  1. Department of Biology, Faculty of Science, University of Shahid Chamran, Ahvaz, Iran

    Fatemeh Sheikhi & Mohammad Roayaei Ardakani

  2. Department of Soil Science, Faculty of Agriculture, University of Shahid Chamran, Ahvaz, Iran

    Naeimeh Enayatizamir

  3. CEIT, Section of Environmental Engineering, Paseo Manuel de Lardizabal 15, 20018, San Sebastián, Spain

    Susana Rodriguez-Couto

  4. IKERBASQUE, Basque Foundation for Science, Alameda de Urquijo 36, 48011, Bilbao, Spain

    Susana Rodriguez-Couto

Authors
  1. Fatemeh Sheikhi
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  2. Mohammad Roayaei Ardakani
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  3. Naeimeh Enayatizamir
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  4. Susana Rodriguez-Couto
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Correspondence to Fatemeh Sheikhi.

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Sheikhi, F., Roayaei Ardakani, M., Enayatizamir, N. et al. The Determination of Assay for Laccase of Bacillus subtilis WPI with Two Classes of Chemical Compounds as Substrates. Indian J Microbiol 52, 701–707 (2012). https://doi.org/10.1007/s12088-012-0298-3

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  • DOI: https://doi.org/10.1007/s12088-012-0298-3

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