Investigation of the Thrombolytic Potential of Exoproteinases Produced by the Micromycete  Tolypocladium inflatum  62A Isolated from the Soils of the White Sea

N. S. Fokichev; Фокичев Н. С.; E. I. Kornienko; Корниенко Е. И.; V. G. Kreyer; Крейер В. Г.; A. A. Osmolovskiy; Осмоловский А. А.

doi:10.31857/S0026364823020071

Investigation of the Thrombolytic Potential of Exoproteinases Produced by the Micromycete Tolypocladium inflatum 62A Isolated from the Soils of the White Sea

Authors: Fokichev N.S.¹, Kornienko E.I.¹, Kreyer V.G.¹, Osmolovskiy A.A.¹
Affiliations:
1. Lomonosov Moscow State University
Issue: Vol 57, No 2 (2023)
Pages: 95-103
Section: ФИЗИОЛОГИЯ, БИОХИМИЯ, БИОТЕХНОЛОГИЯ
URL: https://journals.rcsi.science/0026-3648/article/view/138052
DOI: https://doi.org/10.31857/S0026364823020071
EDN: https://elibrary.ru/NIXIPI
ID: 138052

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Abstract
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Abstract

Thrombolytic properties of the micromycete Tolypocladium inflatum 62a isolated from the White Sea soils exoproteinases preparation, as well as its individual fractions, obtained after isoelectric focusing, were studied in comparison to the closest analogue, the producer of thrombolytic enzymes T. inflatum k1. A pronounced thrombolytic potential, the presence of fibrinolytic and plasminogen activator activity, specific proteolytic activity with respect to certain substrates (plasmin-like and subtilisin-like) as well as a prolonged thrombolytic effect in connection to fibrin clots were demonstrated, which can be used in the development of new drugs for the thrombotic conditions treatment, the development of anti-hematoma drugs for external use as well as diagnostic kits for the pathology of the human hemostasis system.

Keywords

extracellular proteinases, fibrinolytic enzymes, hemostasis system, micromycetes, thrombolysis, thrombotherapy

References

Averina A.V., Snegiryova A.Ya. Laboratory workshop on organic chemistry. Vysshaya shkola, Moscow, 1980 (in Russ.).
Baker W.F. Jr. Thrombolytic therapy. Clin. Appl. Thromb. Hemost. 2002. V. 8 (4). P. 291–314. https://doi.org/10.1177/107602960200800401
Balami J.S., Chen R., Sutherland B.A. et al. Thrombolytic agents for acute ischaemic stroke treatment: the past, present and future. CNS Neurol. Disord. Drug. Targets. 2013. V. 12 (2). P. 145–154. https://doi.org/10.2174/18715273113129990057
Batomunkueva B.P., Egorov N.S. Isolation, purification and resolution of the extracellular proteinase complex of Aspergillus ochraceus 513 with fibrinolytic and anticoagulant activities. Microbiology. 2001. V. 70 (5). P. 519–522 (in Russ.).
Bissett J. Notes on Tolypocladium and related genera. Can. J. Bot. 1983. V. 5. P. 1311–1329. https://doi.org/10.1139/b83-139
Domsch K.H., Gams W., Anderson T.H. Compendium of soil fungi, 2nd ed. IHW-Verlag, Eching, 2007.
Fokichev N.S., Kornienko E.I., Sharkova T.S. et al. Thrombolytic activity and properties of the proteinase produced by the micromycete Tolypocladium inflatum k1. Mikologiya i fitopatologiya. 2021. V. 55. № 6. P. 449–456. https://doi.org/10.31857/S002636482106009X
Hao Q., Dong B.R., Yue J. et al. Thrombolytic therapy for pulmonary embolism. Cochrane Database Syst. Rev. 2018. V. 12. P. 1–105. https://doi.org/10.1002/14651858.CD004437.pub5
Holbrook I.B., Leaver A.G. A procedure to increase the sensitivity of staining by Coomassie Briliant Blue G-250 – perchloric acid solution. Anal. Biohem. 1976. V. 75 (2). P. 634–636.
Khaldi N., Seifuddin F.T., Turner G. et al. SMURF: Genomic mapping of fungal secondary metabolite clusters. FG and B. 2010. V. 47. P. 736–741. https://doi.org/10.1016/j.fgb.2010.06.003
Kotb E., Helal G.E.-D.A., Edries F.M. Screening for fibrinolytic filamentous fungi and enzymatic properties of the most potent producer, Aspergillus brasiliensis AUMC 9735. Biologia. 2015. V. 70 (12). P. 1565–1574. https://doi.org/10.1515/biolog-2015-0192
Osmolovskiy A.A., Kreier V.G., Kurakov A.V. et al. Aspergillus ochraceus micromycetes – producers of extracellular proteinases – protein C activators of blood plasma. Appl. Biochem. Microbiol. 2012. V. 48 (5). P. 488–492.
Osmolovskiy A.A., Kreier V.G., Baranova N.A. et al. Production of extracellular proteinases – protein C activators of blood plasma – by the micromycete Aspergillus ochraceus during submerged and solid-state fermentation. Appl. Biochem. Microbiol. 2013. V. 49 (6). P. 580–586.
Osmolovskiy A.A., Popova E.A., Kreyer V.G. et al. 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 (1). P. 62–66.
Podorolskaya L.V., Serebryakova T.N., Sharkova, T.S. Experimental thrombosis in rabbit marginal ear vein and evaluation of the thrombolytic action of longolytin. Bull. Exp. Biol. Med. 2007. V. 143. P. 577–580. https://doi.org/10.1007/s10517-007-0184-x
Popova E.A., Osmolovskiy A.A., Kreier V.G. et al. Production by Aspergillus ustus strain of proteinases highly activeagainst fibrillar proteins. Mikologiya i fitopatologiya. 2019. V. 53 (4). P. 229–235. https://doi.org/10.1134/S0026364819040111
Seifert K.A., Gams W. The genera of Hyphomycetes–2011 update. Persoonia. 2011. V. 27. P. 1–119. https://doi.org/10.3767/003158511X617435
Sharkova T.S., Kornienko E.I., Osmolovskiy A.A. et al. Morphological and physiological properties of the micromycete Arthrobotrys longa, a producer of longolytin, a proteolytic complex with a thrombolytic effect. Microbiology. 2016a. V. 85 (2). P. 171–176.
Sharkova T.S., Matveeva E.O., Kreier V.G. et al. Production of proteinase – plasminogen activators by micromycete Tolypocladium inflatum k1. Appl. Biochem. Microbiol. 2016b. V. 52 (1). P. 31–35.
Thronton D.J., Carlstadt I., Sheehan J.K. Identification of glycoproteins on nitrocellulose membranes and gels. Mol. Biotechnol. 1996. V. 5 (2). P. 171–176.
White T.J., Bruns T.D., Lee S.B. et al. Amplification and direct sequencing of fungal ribosomal RNA. In: Genes for phylogenetics PCR. Protocols and applications. A laboratory manual, 1st ed. Cambridge, Academic Press, 1990, pp. 315–322.
World Health Organization. 2022. https://www.who.int/data/gho/data/themes/mortality-and-global-health-estimates. Accessed 12.09.2022.
Аверина А.В., Снегирева А.Я. (Averina, Snegireva) Лабораторный практикум по органической химии. М.: Высшая школа, 1980. 184 с.
Батомункуева Б.П., Егоров Н.С. (Batomunkueva, Egorov) Выделение, очистка и разделение комплекса внеклеточных протеиназ Aspergillus ochraceus 513 с фибринолитической и антикоагулянтной активностью // Микробиология. 2001. Т. 70. № 5. С. 519–522.
Осмоловский А.А., Крейер В.Г., Кураков А.В. и др. (Osmolovskiy et al.) Микромицеты Aspergillus ochraceus – продуценты внеклеточных протеиназ – активаторов протеина С плазмы крови // Прикл. биохим. микробиол. 2012. Т. 48. № 5. С. 488–492.
Осмоловский А.А., Крейер В.Г., Баранова Н.А. и др. (Osmolovskiy et al.) Продукция внеклеточных протеиназ – активаторов протеина С плазмы крови – микромицетом Aspergillus ochraceus в ходе глубокой и твердофазной ферментации // Прикл. биохим. микробиол. 2013. Т. 49. № 6. С. 580–586.
Подорольская Л.В., Шаркова Т.С., Андреенко Г.В. и др. (Podorolskaya et al.) Гемостаз и фибринолиз при наружном применении тромболитического препарата лонголитина // Вестник МГУ. Серия биология. 2002. № 2. С. 11–15.
Попова Е.А., Осмоловский А.А., Крейер В.Г. и др. (Popova et al.) Продукция штаммом Aspergillus ustus протеиназ, активных в отношении фибриллярных белков // Микология и фитопатология. 2019. Т. 53. № 4. С. 229–235.
Фокичев Н.С., Корниенко Е.И., Шаркова Т.С. и др. (Fokichev et al.) Тромболитическая активность и свойства препарата протеиназ, образуемых микромицетом Tolypocladium inflatum k1 // Микология и фитопатология. 2021. Т. 55. № 6. С. 449–456.
Шаркова Т.С., Корниенко Е.И., Осмоловский А.А. и др. (Sharkova et al.) Морфофизиологические особенности микромицета Arthrobotrys longa – продуцента протеолитического комплекса тромболитического действия лонголитин // Микробиология. 2016. Т. 85. С. 171–176.
Шаркова Т.С., Матвеева Э.О., Крейер В.Г. и др. (Sharkova et al.) Образование протеиназ – активаторов плазминогена микроскопическим грибом Tolypocladium inflatum k1 // Прикладная биохимия и микробиология. 2016. Т. 52. № 1. С. 31–35.