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Production and Stability of the Proteinase Complex from Aspergillus ochraceus L-1 with Fibrinolytic and Anticoagulant Activity

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Abstract—

A complex of proteolytic enzymes with a specific plasmin-like activity of at least 890 EpNA/mg and a specific protein C activator activity of at least 130 EpNA/mg was isolated from the culture fluid of micromycete Aspergillus ochraceus L-1 by salting out with ammonium sulfate followed by dialysis and depigmentation to phenol-aniline formaldehyde resin at a pH of 8.2. Comparison of the obtained proteinase preparation with caseinolytic, fibrinolytic, fibrinogenolytic, and plasmin-like activities with commercial analogues terrilytin, trypsin, and streptokinase showed its promise for use as a means for thinning purulent burn wounds and fibrin clots. The resulting proteinase complex had high storage stability at low temperatures for up to 9 months.

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REFERENCES

  1. Razzaq, A., Shamsi, S., Ali, A., Ali, Q., Sajjad, M., Malik, A., and Ashraf, M., Microbial proteases applications, Front. Bioeng. Biotechnol., 2019, vol. 7, p. 110.

    Article  PubMed  Google Scholar 

  2. dos Santos Aguilar, J.G. and Sato, H.H., Microbial proteases: Production and application in obtaining protein hydrolysates, Food Res. Int., 2018, vol. 103, pp. 253–262.

    Article  PubMed  Google Scholar 

  3. Kotb, E., The biotechnological potential of fibrinolytic enzymes in the dissolution of endogenous blood thrombi, Biotechnol. Prog., 2014, vol. 30, no. 3, pp. 656–672.

    Article  CAS  PubMed  Google Scholar 

  4. Sharkova, T.S., Kurakov, A.V., Osmolovskiy, A.A., Matveeva, E.O., Kreyer, V.G., Baranova, N.A., and Egorov, N.S., Screening of producers of proteinases with fibrinolytic and collagenolytic activities among micromycetes, Microbiology, 2015, vol. 84, no. 3, pp. 359–364.

    Article  CAS  Google Scholar 

  5. Shamraichuk, I.L., Lavrenova, V.N., Belozerskii, M.A., Kurakov, A.V., Belyakova, G.A., and Dunaevskii, Ya.E., Activity and spectrum of extracellular peptidases in phytopathogenic micromycetes Fusarium anguioides and F. sambucinum,Mikol. Fitopatol., 2016, vol. 50, no. 4, pp. 250–256.

    Google Scholar 

  6. Kotb, E., Helal, G.E.-D.A., and Edries, F.M., Scree-ning for fibrinolytic filamentous fungi and enzymatic properties of the most potent producer, Aspergillus brasiliensis AUMC 9735, Biologia, 2015, vol. 70, no. 12, pp. 1565–1574.

    Article  CAS  Google Scholar 

  7. Osmolovskiy, A.A., Kurakov, A.V., Kreyer, V.G., Baranova, N.A., and Egorov, N.S., Ability of extracellular proteinases of micromycetes Aspergillus flavipes, Aspergillus fumigates, and Aspergillus sydowii to affect proteins of the human haemostatic system, Moscow Univ. Biol. Sci. Bull., 2017, vol. 72, no. 1, pp. 20–24.

    Article  Google Scholar 

  8. Osmolovskiy, A.A., Zvonareva, E.S., Kreyer, V.G., Baranova, N.A., and Egorov, N.S., Secretion of proteinases with fibrinolytic activity by micromycetes of the genus Aspergillus,Moscow Univ. Biol. Sci. Bull., 2018, vol. 73, no. 1, pp. 39–42.

    Article  Google Scholar 

  9. Osmolovskii, A.A., Kreier, V.G., Kurakov, A.V., Baranova, N.A., Piskunkova, N.F., and Egorov, N.S., Micromycetes Aspergillus flavus and A. oryzae as producers of proteinases that activate proteins of the human hemostasis system, Mikol. Fitopatol., 2019, vol. 53, no. 3, pp. 183–185.

    Article  Google Scholar 

  10. Shilpa, H.K., Ambekar, J.G., Dongre, N.N., and Siddalingeshwara, K.G., Application of fibrinolytic enzyme from Aspergillus tamarii – in vitro studies, Eur. J. Pharm. Med. Res., 2019, vol. 6, no. 12, pp. 560–562.

    Google Scholar 

  11. Craik, C.S., Page, M.J., and Madison, E.L., Proteases as therapeutics, Biochem. J., 2011, vol. 435, no. 1, pp. 1–16.

    Article  CAS  PubMed  Google Scholar 

  12. Chanalia, P., Gandhi, D., Jodha, D., and Singh, J., Applications of microbial proteases in pharmaceutical industry: An overview, Rev. Med. Microbiol., 2011, vol. 22, no. 4, pp. 96–101.

    Article  Google Scholar 

  13. Kumar, L. and Jain, S.K., Proteases: A beneficial degradative enzyme in therapeutic applications, Int. J. Sci. Res. Biol. Sci., 2018, vol. 5, no. 4, pp. 114–118.

    Google Scholar 

  14. Osmolovskiy, A.A., Rukavitsyna, E.D., Kreier, V.G., Baranova, N.A., and Egorov, N.S., Production of proteinases with fibrinolytic and fibrinogenolytic activity by a micromycete aspergillus ochraceus, Microbiology, 2017, vol. 86, no. 4, pp. 512–516.

    Article  CAS  Google Scholar 

  15. Osmolovskiy, A.A., Kreyer, V.G., Baranova, N.A., and Egorov, N.S., Properties of extracellular plasmin-like proteases of Aspergillus ochraceus micromycete, Appl. Biochem. Microbiol., 2017, vol. 53, no. 4, pp. 429–434.

    Article  CAS  Google Scholar 

  16. Osmolovskiy, A.A., Orekhova, A.V., Kreyer, V.G., Baranova, N.A., and Egorov, N.S., Possibility of application of extracellular proteases of the micromycete Aspergillus ochraceus VKM F-4104D for determination of the protein C content in human blood plasma, Biochemistry (Moscow)Suppl. Ser. B: Biomed. Chem., 2018, vol. 12, no. 2, pp. 164–166.

    Google Scholar 

  17. Komarevtsev, S.K., Popova, E.A., Kreyer, V.G., Miroshnikov, K.A., and Osmolovskiy, A.A., Purification of the protease activator of protein C of human blood plasma produced by the micromycete Aspergillus ochraceus VKM F-4104D, Appl. Biochem. Microbiol., 2020, vol. 56, no. 1, pp. 39–44.

    Article  Google Scholar 

  18. Shataeva, L.K., Kreier, V.G., Pokrovskaya, S.S., Radzyavichus, K.I., and Samsonov, G.V., Effect of the amino acid composition of the protein on its binding with carboxyl cation exchangers, Vysokomol.Soedin., 1985, vol. 27, no. 1, pp. 46–51.

    CAS  Google Scholar 

  19. Shataeva, L.K., Marina, V.P., Radzyavichus, K.I., Mel’nikova, S.K., and Samsonov, G.V., Effect of swelling of FAF anionite on its sorption properties, Prikl. Biokhim. Mikrobiol., 1982, vol. 18, no. 1, pp. 65–70.

    CAS  Google Scholar 

  20. Batomunkueva, B.P. and Egorov, N.S., Isolation, purification and resolution of the extracellular proteinase complex of Aspergillus ochraceus 513 with fibrinolytic and anticoagulant activities, Microbiology, 2001, vol. 70, no. 5, pp. 519–522.

    Article  CAS  Google Scholar 

  21. Belakhov, V.V. and Momot, N.N., Development of novel sorption method of preparation of high-purified α-amylase for the creation of enzyme drugs, World Scient. Discov., 2010, vol. 6, no. 1, pp. 173–175.

    Google Scholar 

  22. Osmolovskiy, A.A., Kreier, V.G., Kurakov, A.V., Baranova, N.A., and Egorov, N.S., Aspergillus ochraceus micromycetes – producers of extracellular proteinases – protein C activators of blood plasma, Appl. Biochem. Microbiol., 2012, vol. 48, no. 5, pp. 488–492.

    Article  CAS  Google Scholar 

  23. Osmolovskiy, A.A., Popova, E.A., Kreyer, V.G., Baranova, N.A., and Egorov, N.S., Fibrinolytic and collagenolytic activity of extracellular proteinases of the strains of micromycetes Aspergillus ochraceus L-1 and Aspergillus ustus 1, Moscow Univ. Biol. Sci. Bull., 2016, vol. 71, no. 1, pp. 62–66.

    Article  Google Scholar 

  24. Bradford, M.M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem., 1976, vol. 72, nos. 1–2, pp. 248–254.

    Article  CAS  PubMed  Google Scholar 

  25. Lagashetti, A.C., Dufossé, L., Singh, S.K., and Singh, P.N., Fungal pigments and their prospects in different industries, Microorganisms, 2019, vol. 7, no. 12, p. 604.

    Article  CAS  Google Scholar 

  26. Osmolovskiy, A.A., Kreyer, V.G., Baranova, N.A., Kurakov, A.V., and Egorov, N.S., Properties of extracellular proteinase – an activator of protein C in blood plasma formed by Aspergillus ochraceus,Appl. Biochem. Microbiol., 2015, vol. 51, no. 1, pp. 95–101.

    Article  CAS  Google Scholar 

  27. Kotb, E., Purification and partial characterization of serine fibrinolytic enzyme from Bacillus megaterium KSK-07 isolated from Kishk, a traditional Egyptian fermented food, Appl. Biochem. Microbiol., 2015, vol. 51, no. 1, pp. 34–43.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biology, Moscow State University, 119234, Moscow, Russia

    A. A. Osmolovskiy, A. V. Orekhova, V. G. Kreyer & N. A. Baranova

  2. La Sapienza University of Rome, 00185, Rome, Italy

    A. V. Orekhova & E. Conti

  3. International Biotechnological Center, Moscow State University, 119234, Moscow, Russia

    N. S. Egorov

Authors
  1. A. A. Osmolovskiy
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  2. A. V. Orekhova
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  3. E. Conti
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  4. V. G. Kreyer
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  5. N. A. Baranova
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  6. N. S. Egorov
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Correspondence to A. A. Osmolovskiy.

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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.

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Translated by M. Novikova

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Osmolovskiy, A.A., Orekhova, A.V., Conti, E. et al. Production and Stability of the Proteinase Complex from Aspergillus ochraceus L-1 with Fibrinolytic and Anticoagulant Activity. Moscow Univ. Biol.Sci. Bull. 75, 130–135 (2020). https://doi.org/10.3103/S0096392520030074

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  • DOI: https://doi.org/10.3103/S0096392520030074

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