Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Action of Extracellular Protease of Aspergillus terreus on Human Plasma Hemostasis Proteins

  • Alexander A. Osmolovskiy (Department of Microbiology, Faculty of Biology, Lomonosov Moscow State University) ;
  • Elena S. Zvonareva (Higher School of Economics - National Research University) ;
  • Nina A. Baranova (Department of Microbiology, Faculty of Biology, Lomonosov Moscow State University) ;
  • Valeriana G. Kreyer (Department of Microbiology, Faculty of Biology, Lomonosov Moscow State University)
  • Received : 2022.05.06
  • Accepted : 2022.06.30
  • Published : 2023.06.28
Download PDF
⟨ Previous Next ⟩

Abstract

Proteolytic enzymes secreted by Aspergillus, as pathogenicity factors, affect blood coagulation and fibrinolysis, and therefore the target proteins of their action in the bloodstream are of significant interest. In the present study, the action of the isolated protease of A. terreus 2 on different human plasma proteins was shown. The protease of A. terreus 2 exhibited the highest proteolytic activity against hemoglobin, which was 2.5 times higher than the albuminolytic activity shown in both of the protein substrates used. In addition, the protease has significant ability to hydrolyze both fibrin and fibrinogen. However, the inability of the A. terreus 2 protease to coagulate rabbit blood plasma and coagulate human and bovine fibrinogen indicates the severity of the enzyme's action on human blood coagulation factors. It should be considered as a potential indicator of this isolated protease's participation in fungal pathogenesis. The protease shows no hemolytic activity. Furthermore, its activity is insignificantly inhibited by thrombin inhibitors, and is not inhibited by plasmin inhibitors.

Keywords

Acknowledgement

This work was supported by the Financial Council for Grants of the President of the Russian Federation No. SP-3906.2021.4.

References

  1. Yike I. Fungal proteases and their pathophysiological effects. Mycopathologia 171: 299-323.
  2. Ventakatanagaraju E, Divakar G. 2014. An overview on microbial fibrinolytic proteases. Int. J. Pharm. Sci. Res. 5: 643-656.
  3. Sharkova TS, Kurakov AV, Osmolovskiy AA, Matveeva EO, Kreyer VG, Baranova NA, et al. 2015. Screening of producers of proteases with fibrinolytic and collagenolytic activities among micromycete. Microbiology 84: 359-364. https://doi.org/10.1134/S0026261715030182
  4. Timorshina S, Boiko Y, Kornienko E, Orekhova A, Osmolovskiy A. 2021. Fibrinolytic proteases of micromycetes promising in biomedicine and biodegradation. Biotechnol. Biotechnol. Equip. 35: S99-100.
  5. Osmolovskiy AA, Zvonareva ES, Kreyer VG, Baranova NA, Egorov NS. 2014. The effect of micromycete extracellular proteases of the Aspergillus genus on the proteins of haemostatic system. Russ. J. Bioorg. Chem. 40: 634-639. https://doi.org/10.1134/S1068162014060120
  6. Kotb E, Helal GEDA, Edries FM. 2015. Screening for fibrinolytic filamentous fungi and enzymatic properties of the most potent producer, Aspergillus brasiliensis AUMC 9735. Biologia 70: 1565-1574. https://doi.org/10.1515/biolog-2015-0192
  7. Orekhova A, Bednenko D, Osmolovskiy A. 2018. Aspergillus flavus and Aspergillus ochraceus as producers of hemostatically active proteins. Porto. Biomed. J. 3: 10-11.
  8. Osmolovskiy AA, Orekhova AV, Kreyer VG, Baranova NA, Egorov NS. 2018. Possibility of application of extracellular protease of the micromycetе Aspergillus ochraceus VKM F-4104D for determination of the protein C content in human blood plasma. Biochem. Suppl. Ser. B. 12: 164-166. https://doi.org/10.1134/S1990750818020099
  9. Osmolovskiy AA, Zvonareva ES, Kreyer VG, Baranova NA, Egorov NS. 2018. Secretion of proteinases with fibrinolytic activity by micromycetes of the genus Aspergillus. Moscow Univ. Biol. Sci. Bull. 73: 39-42. https://doi.org/10.3103/S0096392518010066
  10. Lukianova AA, Kornienko EI, Vigand PA, Kreyer VG, Kurakov AV, Osmolovskiy AA. 2020. Secretion of proteinase with activity that is similar to activity of proteins of the hemostatic system by micromycetes. Moscow Univ. Biol. Sci. Bull. 75: 37-42. https://doi.org/10.3103/S009639252001006X
  11. Osmolovskiy AA, Lukianova AA, Zvonareva ES, Kreyer VG, Baranova NA, Egorov NS. 2018. Combined microbiological approach to screening of producers of proteases with hemostasis system proteins activity among micromycetes. Biotech. Rep. 19: e00265.
  12. Orekhova AV, Osmolovskiy AA, Kreyer VG, Baranova NA, Egorov NS. 2019. Possibility for application of extracellular protease of micromycete Aspergillus ochraceus for determining factor X content in human blood plasma. Moscow Univ. Biol. Sci. Bull. 74: 117-120. https://doi.org/10.3103/S0096392519020081
  13. Chakrabarti SK, Matsumura N, Ranu RS. 2000. Purification and characterization of an extracellular alkaline serine protease from Aspergillus terreus (IJIRA 6.2). Curr. Microbiol. 40: 239-244. https://doi.org/10.1007/s002849910048
  14. Biaggio RT, Silva RR, Rosa NG, Leite RS, Arantes EC, Cabral T, et al. 2016. Purification and biochemical characterization of an extracellular serine peptidase from Aspergillus terreus. Prep. Biochem. Biotechnol. 46: 298-304. https://doi.org/10.1080/10826068.2015.1031387
  15. de Lima EE, Franco DG, Galeano RMS, Guimaraes NCA, Masui DC, Giannesi GC, et al. 2021. Biochemical characterization of a partially purified protease from Aspergillus terreus 7461 and its application as an environmentally friendly dehairing agent for leather industry. Prep. Biochem. Biotechnol. 51: 320-330. https://doi.org/10.1080/10826068.2020.1815058
  16. Zvonareva ES, Osmolovskiy AA, Kreier VG, Baranova NA, Kotova IB, et al. 2018. Production of proteinase with plasmin-like and prekallikrein activating activity by the micromycete Aspergillus terreus. Appl. Biochem. Microbiol. 54: 206-210. https://doi.org/10.1134/S0003683818020151
  17. Zvonareva ES, Osmolovskiy AA, Kreyer VG, Baranova NA, Kotova IB, et al. 2015. Identification of targets for extracellular proteases activating proteins of the haemostatic system produced by micromycetes Aspergillus ochraceus and Aspergillus terreus. Russ. J. Bioorg. Chem. 41: 500-505. https://doi.org/10.1134/S1068162015050155
  18. Vesterberg O. 1971. Isoelectric focusing of proteins. Methods Enzymol. 22: 389-412. https://doi.org/10.1016/0076-6879(71)22035-8
  19. Davis BJ. 1964. Disc Electrophoresis. II. Method and application to human serum proteins. Ann. N.Y. Acad. Sci. 121: 404-427. https://doi.org/10.1111/j.1749-6632.1964.tb14213.x
  20. Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254. https://doi.org/10.1016/0003-2697(76)90527-3
  21. Osmolovskiy AA, Popova EA, Kreyer VG, Baranova NA, Egorov NS. 2016. 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. 71: 62-66. https://doi.org/10.3103/S0096392516010053
  22. Chung D-M, Choi NS, Maeng PJ, Chun HK, Kim S-H. 2010. Purification and characterization of a novel fibrinolytic enzyme from chive (Allium tuberosum). Food Sci. Biotechnol. 126: 349-355.
  23. Hu Y, Yu D, Wang Z, Hou J, Tyagi R, Liang Y, et al. 2019. Purification and characterization of a novel, highly potent fibrinolytic enzyme from Bacillus subtilis DC27 screened from Douchi, a traditional Chinese fermented soybean food. Sci. Rep. 9: 9235.
  24. Osmolovskiy AA, Schmidt L, Orekhova AV, Komarevtsev SK, Kreyer VG, Shabunin SV, et al. 2021. Action of extracellular proteases of Aspergillus flavus and Aspergillus ochraceus micromycetes on plasma hemostasis proteins. Life 11: 782.