Skip to main content
SpringerLink
Log in

Production of Beluga Whale (Delphinapterus leucas) Chymosin in the Methylotrophic Yeast Komagataella phaffii and Characteristics of the Recombinant Enzyme

  • Published:
Applied Biochemistry and Microbiology Aims and scope Submit manuscript

Abstract

A producer strain of Komagataella phaffii of recombinant beluga whale (Delphinapterus leucas) chymosin with a productivity of 80 mg/L was obtained, and the physico-chemical properties of the enzyme were studied for possible use in the food industry. The specific activity of the recombinant beluga chymosin was 55 IMCU/mg, which is significantly lower than that of recombinant bovine chymosin. The beluga whale chymosin has a high nonspecific proteolytic activity, which impaired its properties and ruled out its used in the food industry. Apparently, the amino acid substitutions K221M and K294Q are critically important and lead to a lower binding of casein micelles to the recombinant chymosin of the beluga whale.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.
Fig. 5.

Similar content being viewed by others

REFERENCES

  1. Kumar, A., Grover, S., Sharma, J., and Batish, V.K., Crit. Rev. Biotechnol., 2010, vol. 30, no. 4, pp. 243–258.

    Article  CAS  Google Scholar 

  2. Kappeler, S.R., Van den Brink, H.(J.)M., Rahbek-Nielsen, H., Farah, Z., Puhan, Z., Hansen, E.B., and Johansen, E., Biochem. Biophys. Res. Commun., 2006, vol. 2, no. 342, pp. 647–654.

    Article  Google Scholar 

  3. Belenkaya, S.V., Balabova, D.V., Belov, A.N., Koval, A.D., Shcherbakov, D.N., and Elchaninov, V.V., Appl. Biochem. Microbiol., 2020, vol. 56, no. 4, pp. 363–372.

    Article  CAS  Google Scholar 

  4. Vallejo, J.A., Ageitos, J.M., Poza, M., and Villa, T.G., J. Dairy Sci., 2012, vol. 95, no. 2, pp. 609–613.

    Article  CAS  Google Scholar 

  5. Ersöz, F. and İnan, M., Protein Expr. Purif., 2019, vol. 154, pp. 126–133.

    Article  Google Scholar 

  6. Vallejo, J.A., Ageitos, J.M., Poza, M., and Villa, T.G., J. Agric. Food Chem., 2008, vol. 56, no. 22, pp. 10606–10610.

    Article  CAS  Google Scholar 

  7. Belenkaya, S.V., Shcherbakov, D.N., Balabova, D.V., Kriger, A.V., Belov, A.N., Koval, A.D., and Elchaninov, V.V., Appl. Biochem. Microbiol., 2018, vol. 54, no. 6, pp. 569–576.

    Article  CAS  Google Scholar 

  8. Belenkaya, S.V., Bondar, A.A., Kurgina, T.A., Elchaninov, V.V., Bakulina, A.Yu., Rukhlova, E.A., Lavrik, O.I., Ilyichev, A.A., and Shcherbakov, D.N., Biochemistry (Moscow), 2020, vol. 85, no. 7, pp. 916–928.

    Google Scholar 

  9. Oftedal, O.T., in Encyclopedia of Dairy Sciences, Fuquay, J., Fox, P.F., and McSweeney, P., Eds., San Diego, CA: Academic, 2011, vol. 3, pp. 563–580.

    Google Scholar 

  10. Manguy, J. and Shields, D.C., R. Soc. Open Sci., 2019, vol. 6, no. 10, pp. 1–17.

    Article  Google Scholar 

  11. Noseda, D.G., Recupero, M., Blasco, M., Bozzo, J., and Galvagno, M.A., Protein Expr. Purif., 2016, vol. 123, pp. 112–121.

    Article  CAS  Google Scholar 

  12. Fil’kin, S.Yu., Chertova, N.V., Vavilova, E.A., Zatsepin, S.S., El’darov, M.A., Sadykhov, E.G., Fedorov, A.N., and Lipkin, A.V., Appl. Biochem. Microbiol., 2020, vol. 56, no. 6, pp. 657–661.

    Article  Google Scholar 

  13. Wang, N., Wang, K.Y., Li, G., Guo, W., and Liu, D., Protein Expr. Purif., 2015, vol. 111, pp. 75–81.

    Article  CAS  Google Scholar 

  14. Sagar, A.J. and Pandit, M.W., Biochim. Biophys. Acta, 1983, vol. 743, no. 3, pp. 303–309.

    Article  CAS  Google Scholar 

  15. Laemmli, U.K., Nature, 1970, vol. 227, no. 5259, pp. 680–685.

    Article  CAS  Google Scholar 

  16. Langholm, JensenJ., Molgaard, A., Navarro, Poulsen, J.C., Harboe, M.K., Simonsen, J.B., Lorentzen, A.M., Hjerno, K., Brink, J.M., Qvist, K.B., and Larsen, S., Acta Crystallogr. D Biol. Crystallogr., 2013, vol. 69, no. 5, pp. 901–913.

    Article  Google Scholar 

  17. Higgins, D.R., Busser, K., Comiskey, J., Whittier, P.S., Purcell, T.J., and Hoeffler, J.P., Methods Mol. Biol., 1998, vol. 103, pp. 41–53.

    CAS  PubMed  Google Scholar 

Download references

ACKNOWLEDGMENTS

This study was performed at the facilities of the Industrial Biotechnologies Center for Collective Use of the Fundamentals of Biotechnology Federal Research Center of the Russian Academy of Sciences.

Funding

This study was supported by the Ministry of Science and Higher Education of the Russian Federation, the agreement of November 26, 2018, No. 14.607.21.0207 (unique identifier RFMEFI60718X0207), the Federal Target Program “Research and Development in Priority Areas of the Scientific-Technological Complex of Russia for 2014–2020.”

Author information

Authors and Affiliations

  1. Bach Institute of Biochemistry, Federal Research Center Fundamentals of Biotechnology, Russian Academy of Sciences, 119071, Moscow, Russia

    S. Y. Filkin, N. V. Chertova, S. S. Zatsepin, E. G. Sadykhov, A. N. Fedorov & A. V. Lipkin

Authors
  1. S. Y. Filkin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. N. V. Chertova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. S. Zatsepin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. E. G. Sadykhov
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A. N. Fedorov
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A. V. Lipkin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A. V. Lipkin.

Ethics declarations

The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

Additional information

Translated by M. Novikova

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Filkin, S.Y., Chertova, N.V., Zatsepin, S.S. et al. Production of Beluga Whale (Delphinapterus leucas) Chymosin in the Methylotrophic Yeast Komagataella phaffii and Characteristics of the Recombinant Enzyme. Appl Biochem Microbiol 57, 297–302 (2021). https://doi.org/10.1134/S0003683821030029

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0003683821030029

Keywords:

Search

Navigation