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Production of Maral (Cervus elaphus sibiricus Severtzov) Recombinant Chymosin in the Prokaryotic Expression System and the Study of the Aggregate of Its Biochemical Properties Relevant for the Cheese-Making Industry

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Abstract

The paper reports on the production of the maral recombinant chymosin in the Escherichia coli expression system (SHaffle express strain) and the study of its biochemical properties relevant for the cheese-making industry. The highest maral recombinant prochymosin content in inclusion bodies was observed when producer cells were cultured at 25°C for 6 h after the introduction of an inducer, 10 mM isopropyl-β-D-1-thiogalactopyranoside. The biochemical properties of the obtained enzyme were compared with those of bovine and dromedarian recombinant chymosins. It is shown that total proteolytic activity of the maral recombinant chymosin was comparable with that of the bovine enzyme and that it exceeded the activity of the dromedarian enzyme by about 3.8 times. The thermal stability of the recombinant chymosin from maral was found to be 5–10°C higher than that of the chymosins from the cow and dromedary. The patterns of dependence of milk-clotting activity on the pH level and calcium chloride concentration in cow’s milk on the enzyme from maral met the requirements set by the cheese industry. Its high proteolytic activity and thermal stability limits the scope of application of maral recombinant chymosin to the production of cheeses with short ripening and short storage times.

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Funding

The work was supported by the State Task of the Ministry of Science and Education of the Russian Federation (topic no. FZMW-2020-0002, “Design of recombinant enzyme producers for the cheese-making industry”).

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Authors and Affiliations

  1. Vector State Scientific Center of Virology and Biotechnology, 630559, Kol’tsovo, Russia

    S. V. Belenkaya & D. N. Shcherbakov

  2. Novosibirsk State University, 630090, Novosibirsk, Russia

    S. V. Belenkaya

  3. Altai State University, 656049, Barnaul, Russia

    D. N. Shcherbakov & D. V. Balabova

  4. Federal Altai Scientific Center of Agrobiotechnologies, Siberian Research Institute of Cheese-Making, 656016, Barnaul, Russia

    A. N. Belov, A. D. Koval & V. V. Elchaninov

Authors
  1. S. V. Belenkaya
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  2. D. N. Shcherbakov
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  3. D. V. Balabova
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  4. A. N. Belov
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  5. A. D. Koval
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  6. V. V. Elchaninov
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Corresponding author

Correspondence to V. V. Elchaninov.

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The authors declare that they have no conflict of interest. The paper contains no experimentation involving animals or humans performed by any of the authors.

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Translated by E. Martynova

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Belenkaya, S.V., Shcherbakov, D.N., Balabova, D.V. et al. Production of Maral (Cervus elaphus sibiricus Severtzov) Recombinant Chymosin in the Prokaryotic Expression System and the Study of the Aggregate of Its Biochemical Properties Relevant for the Cheese-Making Industry. Appl Biochem Microbiol 56, 647–656 (2020). https://doi.org/10.1134/S0003683820060034

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  • DOI: https://doi.org/10.1134/S0003683820060034

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