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

  • Mahnaz Mohtashami
  • Jamshid Fooladi
  • Aliakbar Haddad-Mashadrizeh
  • Mohammadreza Housaindokht
Original Article
  • 46 Downloads

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.

Keywords

Laccases POXA1b Codon optimization Cloning and expression ABTS Benzo[a]pyrene 

Notes

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.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Mahnaz Mohtashami
    • 1
  • Jamshid Fooladi
    • 2
  • Aliakbar Haddad-Mashadrizeh
    • 3
    • 4
  • Mohammadreza Housaindokht
    • 5
  1. 1.Department of Microbiology, Faculty of Biological ScienceAlzahra UniversityTehranIran
  2. 2.Department of Biotechnology, Faculty of Biological ScienceAlzahra UniversityTehranIran
  3. 3.Recombinant Proteins Research Group, Institute of BiotechnologyFerdowsi University of MashhadMashhadIran
  4. 4.Department of Biology, Faculty of SciencesFerdowsi University of MashhadMashhadIran
  5. 5.Department of Chemistry, Faculty of SciencesFerdowsi University of MashhadMashhadIran

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