Screening of producers of proteolytic enzymes effective against fibrillar and globular proteins, among the microfungi associated with insects

Capa

Citar

Texto integral

Acesso aberto Acesso aberto
Acesso é fechado Acesso está concedido
Acesso é fechado Somente assinantes

Resumo

Thirty cultures of microfungi associated with insects were isolated and identified. These cultures were used to study the formation of proteolytic enzymes by the injection seeding method on agarized nutrient media containing sources of specific fibrillar and globular protein substrates. Data was collected on the presence of caseinolytic (EI from 1.00 to 2.75), collagenolytic (EI from 1.00 to 3.25), gelatinolytic (EI from 1.00 to 1.84), elastolytic (EI from 1.00 to 1.50), keratinolytic (EI from 1.00 to 2.00), and hemoglobin hydrolysis abilities (EI from 1.00 to 1.67) for promising strains. Promising strains were identified that showed high activity against tested substrates – nos 12 (Penicillium hirsutum 1), 19 (Cladosporium sphaerospermum 1), 21 (Aspergillus unguis 1), and 25 (Cladosporium herbarum 1). Submerged cultivation of these promising strains was carried out, and the proteolytic activity of extracellular proteases in the culture liquid was determined in relation to various protein substrates. The highest level of total proteolytic activity was determined in the culture liquid of Aspergillus ochraceus 1 (Eazocasein = 1.686 con.units/ml), collagenolytic activity during cultivation of the strain Cladosporium herbarum 1 (Eazocoll = 0.110 con. units/ml), as well as elastolytic activity using the chromogenic peptide substrate S-4760 in the culture fluid of the micromycete Aspergillus unguis 2 (ES-4760 = 7.366 mmol pNA × 10–3/ml/min) under conditions of deep cultivation of insect-associated microfungi strains.

Texto integral

Acesso é fechado

Sobre autores

D. Basalaeva

Lomonosov Moscow State University

Autor responsável pela correspondência
Email: dbasalaewa@yandex.ru
Rússia, Moscow

A. Bogomolova

Utrecht University

Email: a1a2bogomolova@gmail.com
Rússia, Utrecht

A. Osmolovsky

Lomonosov Moscow State University

Email: aosmol@mail.ru
Rússia, Moscow

A. Aleksandrova

Lomonosov Moscow State University

Email: alina-alex2011@yandex.ru
Rússia, Moscow

Bibliografia

  1. Ballester A.-R., Lopez-Perez M., de la Fuente B. et al. Functional and pharmacological analyses of the role of Penicillium digitatum proteases on virulence. Microorganisms. 2019. V. 7 (7). P. 198–217. https://doi.org/10.3390/microorganisms7070198
  2. Barrett A.J. Proteolytic enzymes: serine and cystein peptidases. Methods in Enzymology. 1994. V. 244. V. 244 (1). P. 1–15. https://doi.org/10.1006/abio.1995.1306
  3. Bednenko D.M., Kreyer V.G., Baranova N.A. et al. Biotechnological potential of proteolytic enzymes of micromycete Aspergillus flavus O-1. Advances in medical mycology. 2018. V. 19. P. 108–110. (In Russ.) https://doi.org/10.14427/amm.2018.xix.08
  4. Braunstein A.E. Nomenclature of enzymes: Recommendations of the International Biochemical Union on the nomenclature and classification of enzymes, symbols of the kinetics of enzymatic reactions. VINITI, Moscow, 1979. (In Russ.)
  5. Bukharin O.V. (ed.). Associative symbiosis. Publishing House of the Ural Branch of the Russian Academy of Sciences, Ekaterinburg, 2007. (In Russ.)
  6. Castellanos-Moguel J., Gonzales-Barajas M., Mier T. et al. Virulence testing and extracellular subtilin-like (Pr1) and trypsin-like (Pr2) activity during propagule production of Paecilomyces fumosoroseus isolated from whiteflies (Homoptera: Aleyrodidae). Revista Iberoamericana de Micología. 2007. V. 24 (1). P. 62–68. https://doi.org/10.1016/s1130-1406(07)70016-5
  7. Douglas A.E. Symbiotic interaction. Oxford Univ. Press, Toronto, 1994.
  8. Emtsev V.T. Associative symbiosis of soil diazotrophic bacteria and vegetable crops. Pochvovedenie. 1994. No. 4. P. 74– 84. (In Russ.)
  9. Evlakhova A.A. Entomopatogenny`e griby`. Sistematika, biologiya, prakticheskoe znachenie. Leningrad: Nauka, 1974. 260 s.
  10. Fukuda R., Horiuchi F., Ohta A. et al. The prosequence of Rhizopus niveus asparatic proteinase-I supports correct folding and secretion of its mature part in Saccharomyces cerevisiae. J. Biol. Chem. 1994. V. 269 (13). P. 9556–9561.
  11. Ganbarov H.G., Safarova A. Kh., Shafieva S.M. Proteolytic activity of fungi of the genus Aspergillus isolated from the soil of Azerbaijan. Izvestiya Ufimskogo nauchnogo tsentra RAN. 2018. V. 3 (1). P. 80–84. (In Russ.) https://doi.org/10.31040/2222-8349-2018-1-3-80-84
  12. Giovannoni M., Larini I., Scafati V. A novel Penicillium sumatranse isolate reveals an arsenal of degrading enzymes exploitable in algal biorefinery processes. Biotechnol. Biofuels Bioproducts. 2021. V. 14 (1). P. 180–199. https://doi.org/10.1186/s13068-021-02030-9
  13. Gzogyan L.A. Proteolytic enzymes and their inhibitors in grown mushrooms. Cand. Sci. Thesis. Krasnodar, 2005. (In Russ.)
  14. Lacey L.A., Kaya K.H. Field manual of techniques in invertebrate pathology: application and evaluation of pathogens for control of insects and other invertebrate pests. Springer, Davis, 2007.
  15. Lui S.Q., Meng Z.H., Jang J.K. et al. Characterizing structural features of cuticle-degrading proteases from fungi by molecular modeling. BMC Struct. Biol. 2007. N7. A 33. https://doi.org/10.1186/1472-6807-7-33
  16. Lukin S.A., Kevin P.A., Zvyagintsev D. Yu. Azospirilli and associative nitrogen fixation in non-legume crops in agricultural practice. Selskokhozyaystvennaya biologiya. 1987. N1. P. 51–58. (In Russ.)
  17. Lysenko L.A., Nemova N.N., Kancerova I.P. Proteolytic regulation of biological processes. Karelian Scientific Center of the Russian Academy of Sciences, Petrozavodsk, 2011. (In Russ.)
  18. Osmolovskiy A.A., Popova E.A., Kreyer V.G. et al. Fibrinolytic and collagenolytic activity of extracellular proteinases of micromycete strains Aspergillus ochraceus l-1 and Aspergillus ustus 1. Vestnik Moskovskogo universiteta. Seriya 16. Biologiya. 2016. N1. P. 71–75. (In Russ.) https://doi.org/10.3103/S0096392516010053
  19. Popova E.A., Osmolovskiy A.A., Kreyer V.G. et al. Production of proteinases highly active against fibrillar proteins by Aspergillus ustus strain. Mikologiya i fitopatologiya. 2019. V. 53 (4). P. 229–235. (In Russ.) https://doi.org/10.1134/S0026364819040111
  20. Provorov N.A. Genetic and its evolutionary basis of the theory of symbiosis. Zhurnal obshchey biologii. 2001. V. 62. P. 472–495. (In Russ.) http://doi.org/10.1134/C0026364819020089
  21. Rimareva L.V., Overchenko M.B., Morozova K.A. Patent 2315096 C1 Russian Federation No. of the fungus Aspergillus oryzae, producing a complex of proteinases, pectinases, β-glucanases, α-amylase and xylanase. Application No. 2006123552/13 dated 04.07.2006; published 20.01.2008, Bulletin N2. (In Russ.)
  22. Semenova T.A., Dunaevsky Ya.E., Belyakova G.A. et al. Extracellular peptidases of insect-associated fungi and their possible use in biological control programs and as pathogenicity markers. Fungal Biol. 2020. V. 1 (124). P. 65–72. https://doi.org/10.1016/j.funbio.2019.11.005
  23. Semenova T.A. Extracellular peptidases of fungi forming biotic connections with insects. Cand. Biol. Thesis Abstract. Moscow, 2001. (In Russ.)
  24. Shamraichuk I.L., Belyakova G.A., Eremina I.M. et al. Proteolytic enzymes of fungi and their inhibitors as promising and biocidal agents with antifungal action. Vestnik Moskovskogo universiteta. 2020. V. 75 (3). P. 123–130.
  25. Sharma H., Rai A.K., Chettri R. et al. Bioactivities of Penicillium citrinum isolated from a medicinal plant Swertia chirayita. Arch. Microbiol. 2021. V. 203(8). P. 5173–5182. https://doi.org/10.1007/s00203-021-02498-x
  26. Soltan M.A., Eldeen M.A., Elbassiouni N. et al. In silico designing of a multitope vaccine against Rhizopus microspores with potential activity against other mucormycosis causing fungi. Cells. 2021. V. 10 (11). P. 1–25. https://doi.org/10.3390/cells10113014
  27. Veiter L., Rajamanickam V., Herwig C. The filamentous fungal pellet-relationship between morphology and productivity. Appl. Microbiol. Biotechnol. 2018. V. 102. P. 2997–3006. https://doi.org/10.1007/s00253-018-8818-7
  28. Velikoreczkaya I.A. Creation of new complex farmer’s preparations of fungal proteins based on Penicillium canescens for effective conversion of protein-containing plant serum. Cand. Tech. Thesis. Moscow, 2019. (In Russ.)
  29. Yakhyaeva M.A., Akhmedova Z.R. Some properties of proteolytic enzymes of the fungus Aspergillus oryzae-5. Universum: technical sciences. 2020. N9–2 (78). P. 50–54. (In Russ.)
  30. Ye F., Liang L., Mi Q. et al. Preliminary crystallographic study of two cuticle-degrading proteases from the nematophagous fungi Lecanicillium psalliotae and Paecilomyces lilacinus. Acta Crystallogr. Sect. F: structural biology and crystallization communications. 2009. V. 1 (65). P. 271–274. https://doi.org/10.1107/S1744309109003595
  31. Бедненко Д.М., Крейер В.Г., Баранова Н.А. и др. (Bednenko et al.) Биотехнологический потенциал протеолитических ферментов микромицета Aspergillus flavus O-1 // Успехи медицинской микологии. 2018. Т. 19. С. 108–110.
  32. Браунштейн А.Э. (Braunstein) Номенклатура ферментов: Рекомендации Международного биохимического союза по номенклатуре и классификации ферментов, символам кинетики ферментативных реакций. М.: ВИНИТИ, 1979. 322 с.
  33. Бухарин О.В. (ред.). (Bukharin) Ассоциативный симбиоз. Екатеринбург: Изд-во УрО РАН, 2007. 265 c.
  34. Великорецкая И.А. (Velikoretskaya) Создание новых комплексных агропрепаратов грибных белков на основе Penicillium canescens для эффективной переработки белоксодержащей растительной сыворотки. Дисс. … канд. техн. наук. Москва, 2019. 151 с.
  35. Ганбаров Х.Г., Сафарова А.Х., Шафиева С.М. (Ganbarov et al.) Протеолитическая активность грибов рода Aspergillus, выделенных из почвы Азербайджана // Известия Уфимского научного центра РАН. 2018. № 3 (1). С. 80–84.
  36. Гзогян Л.А. (Gzoghyan) Протеолитические ферменты и их ингибиторы в культивируемых грибах. Диссертация на соискание ученой степени кандидата биологических наук. Краснодар, 2005. 20 c.
  37. Емцев В.Т. (Emtsev) Ассоциативный симбиоз почвенных диазотрофных бактерий и овощных культур. Почвоведение. 1994. № 4. С. 74–84.
  38. Лукин С.А., Кевин П.А. Звягинцев Д.Ю. (Lukin et al.) Азоспириллы и ассоциативная азотфиксация у небобовых культур в практике сельского хозяйства // Сельхохозяйственная биология. 1987. № 1. С. 51–58.
  39. Лысенко Л.А., Немова Н.Н., Канцерова И.П. (Lysenko et al.) Протеолитическая регуляция биологических процессов. Петрозаводск: Карельский научный центр РАН, 2011. 482 c.
  40. Осмоловский А.А., Попова Е.А., Крейер В.Г. и др. (Osmolovsky et al.) Фибринолитическая и коллагенолитическая активность внеклеточных протеиназ штаммов микромицетов Aspergillus ochraceus l-1 и Aspergillus ustus 1 // Вестн. Московского ун-та. Сер. 16. Биол. 2016. № 1. С. 71–75.
  41. Попова Е.А., Осмоловский А.А., Крейер В.Г. и др. (Popova et al.) Продукция штаммом Aspergillus ustus протеиназ, высокоактивных в отношении фибриллярных белков // Микология и фитопатология. 2019. Т. 53. № 4. С. 229–235.
  42. Проворов Н.А. (Provorov) Генетика и эволюционные основы теории симбиоза // Журнал общей биологии. 2001. Т. 62. С. 472–495.
  43. Римарева Л.В., Оверченко М.Б., Морозова К.А. (Rimareva et al.) Патент 2315096 С1 Российская Федерация № гриба Aspergillus oryzae, продуцирующий комплекс протеиназ, пектиназ, β-глюканазы, α-амилазы и ксиланазы. Заявка № 2006123552/13 от 04.07.2006; опубл. 20.01.2008, Бюл. № 2.
  44. Семенова Т.А. (Semenova) Внеклеточные пептидазы грибов, образующих биотические связи с насекомыми. Автореферат дисс. … канд. биол. наук. М., 2001. 26 с.
  45. Шамрайчук И.Л., Белякова Г.А., Еремина И.М. и др. (Shamraychuk et al.) Протеолитические ферменты грибов и иx ингибиторы как перспективные и биоцидные средства антифунгального действия. Вестник Московского университета. 2020. Т. 75, № 3. С. 123–130.
  46. Яхьяева М.А. Ахмедова З.Р. (Yakhyaeva, Akhmedova) Некоторые свойства протеолитических ферментов гриба Aspergillus oryzae-5. Universum: технические науки. 2020. № 9–2 (78). С. 50–54.

Arquivos suplementares

Arquivos suplementares
Ação
1. JATS XML
Baixar
2. Fig. 1. Photographs of the studied strains on Sabouraud medium: 1 – strain 30 (Aspergillus carbonarius 1); 2 – strain 21 (A. unguis 2); 3 – strain 25 (Cladosporium herbarum 1); 4 – strain 27 (Mucor plumbeus 1); 5 – strain 15 (M. circinelloides 1); 6 – strain 12 (Penicillium hirsutum 1); 7 – strain 26 (P. aurantiogriseum 2); 8 – strain 24 (Ulocladium botrytis 1).

Baixar (348KB)
Metadados

Declaração de direitos autorais © Russian Academy of Sciences, 2025