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Molecular cloning, expression and characterization of poxa1b gene from Pleurotus ostreatus

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Abstract

In recent decades, fungus laccases (p-diphenol-dioxygen oxidoreductases; EC 1.10.3.2) have attracted the attention of researches due to their wide range of biotechnological and industrial applications. In the present study, we have cloned a gene encoding laccase (poxa1b) from Pleurotus ostreatus and then heterologously expressed in Escherichia coli BL21. The biochemical properties of POXA1b were characterized using ABTS as a typical substrate of laccases. Moreover, the in vitro oxidation of the benzo[a]pyrene was investigated in the presence or absence of ABTS. The codon-optimized poxa1b showed higher expression yields and efficiency in comparison with the wild-type (p < 0.01). The maximum activity of POXA1b (2075 UL-1) was observed after incubation at 50 °C for 0.5 h and the enzyme retained more than 85% of its initial activity after 2 h incubation at 25–45 °C. The optimum pH of the enzyme was pH4 and the enzyme was stable when being incubated at pH range from 2.5 to 4.5 for 2 h in the absence of ABTS, the enzyme oxidized a little amount of benzo[a]pyrene, whereas its oxidation enhanced following the ABTS addition. These findings indicate POXA1b of P. ostreatus as a promising candidate for further biotechnological approaches.

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Acknowledgements

The authors wish to thank Khayam Bioeconomy Institute (KBI) and Alzahra University for supporting this work. This publication represents a section of one PhD thesis by Ms. Mahnaz Mohtashami at the faculty of Biological Science of Alzahra University, Tehran, Iran.

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Authors and Affiliations

  1. Department of Microbiology, Faculty of Biological Science, Alzahra University, Tehran, Iran

    Mahnaz Mohtashami

  2. Department of Biotechnology, Faculty of Biological Science, Alzahra University, Tehran, Iran

    Jamshid Fooladi

  3. Recombinant Proteins Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

    Aliakbar Haddad-Mashadrizeh

  4. Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran

    Aliakbar Haddad-Mashadrizeh

  5. Department of Chemistry, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran

    Mohammadreza Housaindokht

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  1. Mahnaz Mohtashami
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Mohtashami, M., Fooladi, J., Haddad-Mashadrizeh, A. et al. Molecular cloning, expression and characterization of poxa1b gene from Pleurotus ostreatus. Mol Biol Rep 46, 981–990 (2019). https://doi.org/10.1007/s11033-018-4555-3

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