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Gene cloning and characterization of a novel recombinant 40-kDa heat shock protein from Mesobacillus persicus B48

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Abstract

The present study reports the recognition and characterization of the gene encoding the co-chaperone DnaJ in the halophilic strain Mesobacillus persicus B48. The new extracted gene was sequenced and cloned in E. coli, followed by protein purification using a C-terminal His-tag. The stability and function of the recombinant DnaJ protein under salt and pH stress conditions were evaluated. SDS-PAGE revealed a band on nearly 40-kDa region. The homology model structure of new DnaJ demonstrated 56% similarity to the same protein from Streptococcus pneumonia. Fluorescence spectra indicated several hydrophobic residues located on the protein surface, which is consistent with the misfolded polypeptide recognition function of DnaJ. Spectroscopic results showed 56% higher carbonic anhydrase activity in the presence of the recombinant DnaJ homolog compared to its absence. In addition, salt resistance experiments showed that the survival of recombinant E. coli+DnaJ was 2.1 times more than control cells in 0.5 M NaCl. Furthermore, the number of recombinant E. coli BL21+DnaJ colonies was 7.7 times that of the control colonies in pH 8.5. Based on the results, DnaJ from the M. persicus can potentially be employed for improving the functional features of enzymes and other proteins in various applications.

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Acknowledgements

The authors thank the Research Council of University of Guilan for the financial support to this study.

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Authors and Affiliations

  1. Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran

    Fatemeh Fathinejad, Hossein Ghafouri & Arghavan Alizadeh

  2. Medical Biology Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran

    Ebrahim Barzegari

  3. Molecular Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran

    Sajjad Sarikhan

  4. Nuffield Department of Surgical Science, University of Oxford, Oxford, UK

    Newton Howard

  5. Department of Marine Sciences, The Caspian Sea Basin Research Center, University of Guilan, Rasht, Iran

    Hossein Ghafouri

Authors
  1. Fatemeh Fathinejad
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  2. Hossein Ghafouri
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  3. Ebrahim Barzegari
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  4. Sajjad Sarikhan
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  5. Arghavan Alizadeh
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  6. Newton Howard
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Contributions

FF: Conducted experiments and wrote the manuscript and analyzed data. HG:Conceived and designed the research. Project funding and resources were acquired by HG and SS. EB: Analyzed data and contributed new analytical tools. NH Performed the analysis. All authors read and approved the manuscript.

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Correspondence to Hossein Ghafouri.

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Fathinejad, F., Ghafouri, H., Barzegari, E. et al. Gene cloning and characterization of a novel recombinant 40-kDa heat shock protein from Mesobacillus persicus B48. World J Microbiol Biotechnol 39, 248 (2023). https://doi.org/10.1007/s11274-023-03693-2

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