Rational Design and Mutagenesis of Fungal Luciferase from Neonothopanus nambi


The recently described bioluminescent system from fungi has great potential for developing highly efficient tools for biomedical research. Luciferase enzyme is one of the most crucial components of this system. The luciferase from Neonothopanus nambi fungus belongs to the novel still undescribed protein family. The structure data for this protein is almost absent. A detailed study of the N. nambi luciferase properties is necessary for the improvement of analytical methods based on the fungal bioluminescent system. Here we present the positions of key amino acid residues and their effect on enzyme function described using bioinformatic and experimental approaches. These results are useful for further fungal luciferase structure determination.

This is a preview of subscription content, access via your institution.

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.


  1. 1

    Yan, Y., Shi, P., Song, W., and Bi, S., Chemiluminescence and bioluminescence imaging for biosensing and therapy: in vitro and in vivo perspectives, Theranostics, 2019, vol. 9, pp. 4047–4065. https://doi.org/10.7150/thno.33228

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  2. 2

    Fleiss, A. and Sarkisyan, K.S., A brief review of bioluminescent systems, Curr. Genet., 2019, vol. 65, pp. 877–882. https://doi.org/10.1007/s00294-019-00951-5

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  3. 3

    Pomper, M.G. and Gelovani, J.G., Molecular Imaging in Oncology, CRC Press, 2008.

    Book  Google Scholar 

  4. 4

    Chew, A.L.C., Desjardin, D.E., Tan, Y.-S., et al., Bioluminescent fungi from Peninsular Malaysia—a taxonomic and phylogenetic overview, Fungal Diversity, 2015, vol. 70, pp. 149–187. https://doi.org/10.1007/s13225-014-0302-9

    Article  Google Scholar 

  5. 5

    Shimomura, O., Bioluminescence: Chemical Principles and Methods, Singapore: World Scientific Publishing, 2006. https://doi.org/10.1142/6102

    Book  Google Scholar 

  6. 6

    Kotlobay, A.A., Sarkisyan, K.S., Mokrushina, Y.A., et al., Genetically encodable bioluminescent system from fungi, Proc. Natl. Acad. Sci. U. S. A., 2018, vol. 115, pp. 12728–12732. https://doi.org/10.1073/pnas.1803615115

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. 7

    Mitiouchkina, T., Mishin, A.S., Somermeyer, L.G., et al., Plants with genetically encoded autoluminescence, Nat. Biotechnol., 2020. https://doi.org/10.1038/s41587-020-0500-9

  8. 8

    Oliveira, A.G., Desjardin, D.E., Perry, B.A., and Stevani, C.V., Evidence that a single bioluminescent system is shared by all known bioluminescent fungal lineages, Photochem. Photobiol. Sci., 2012, vol. 11, pp. 848–852. https://doi.org/10.1039/c2pp25032b

    CAS  Article  PubMed  Google Scholar 

  9. 9

    Kaskova, Z.M., Dörr, F.A., Petushkov, V.N., et al., Mechanism and color modulation of fungal bioluminescence, Sci. Adv., 2017, p. e1602847. https://doi.org/10.1126/sciadv.1602847

  10. 10

    Bok, J.W. and Keller, N.P., Fast and easy method for construction of plasmid vectors using modified quick-change mutagenesis, Methods Biochem. Analysis, 2012, vol. 944, pp. 163–174. https://doi.org/10.1007/978-1-62703-122-6_11

    CAS  Article  Google Scholar 

  11. 11

    Pichia expression kit. A manual of methods for expression of recombinant proteins using pPICZ and PICZ-alpha, in Pichia pastoris Catalog, 2009. http://tools.thermofisher.com/content/sfs/manuals/pich_man.pdf.

  12. 12

    Kwolek-Mirek, M. and Zadrag-Tecza, R., Comparison of methods used for assessing the viability and vitality of yeast cells, FEMS Yeast Res., 2014, vol. 14, pp. 1068–1079. https://doi.org/10.1111/1567-1364.12202

    CAS  Article  PubMed  Google Scholar 

Download references


This work was supported by the grant from the Russian Science Foundation no. 16-14-00052P, alanine screening was performed by the President grant for leading scientific schools NSh-2605.2020.4.

Author information



Corresponding author

Correspondence to K. V. Purtov.

Ethics declarations


This article does not contain any studies involving animals or human participants performed by any of the authors.


The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Beregovaya, K.A., Myshkina, N.M., Chepurnykh, T.V. et al. Rational Design and Mutagenesis of Fungal Luciferase from Neonothopanus nambi . Dokl Biochem Biophys 496, 14–17 (2021). https://doi.org/10.1134/S1607672921010026

Download citation


  • bioluminescence
  • luciferase
  • Neonothopanus nambi
  • rational design