New micromycetes strains with keratinolytic activity

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Abstract

Abstract. The ability of the micromycete Tolypocladium inflatum to synthesize keratinolytic enzymes has been demonstrated for the first time. 22 strains of micromycetes isolated from enrichment cultures of keratinolytic microorganisms belonged to the genera Aspergillus, Cladosporium, Fusarium, Keratinophyton, Penicillium, Pseudallscheria, Tolypocladium and Trichoderma. Only two strains showed high keratinolytic activity during submerged fermentation – Keratinophyton terreum C106 (74.2 E) and Tolypocladium inflatum ST1 (87.1 E). The target activity of K. terreum C106 in submerged culture decreased almost to zero by the 7th day of cultivation, while the activity of T. inflatum ST1 decreased by less than 20%. An extracellular protease with keratinolytic activity from the micromycete T. inflatum ST1 with a pI of about 5.6 and a molecular weight of about 31 kDa was isolated.

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About the authors

S. N. Timorshina

Lomonosov Moscow State University

Author for correspondence.
Email: timorshina.svetlana@mail.ru
Russian Federation, Moscow, 119234

E. A. Ganetskaya

Lomonosov Moscow State University

Email: timorshina.svetlana@mail.ru
Russian Federation, Moscow, 119234

A. A. Shestakova

Lomonosov Moscow State University; National Research University “Higher School of Economics”

Email: timorshina.svetlana@mail.ru
Russian Federation, Moscow, 119234; Moscow, 101000

V. M. Lyamina

Lomonosov Moscow State University

Email: timorshina.svetlana@mail.ru
Russian Federation, Moscow, 119234

A. V. Aleksandrova

Lomonosov Moscow State University

Email: timorshina.svetlana@mail.ru
Russian Federation, Moscow, 119234

E. A. Popova

Lomonosov Moscow State University

Email: timorshina.svetlana@mail.ru
Russian Federation, Moscow, 119234

G. B. Admanova

Aktobe Regional University named after K. Zhubanova

Email: timorshina.svetlana@mail.ru
Kazakhstan, Aktobe, 030000

A. A. Osmolovsky

Lomonosov Moscow State University

Email: timorshina.svetlana@mail.ru
Russian Federation, Moscow, 119234

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Supplementary files

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2. Fig. 1. Enzymatic indices of micromycete strains isolated from enrichment cultures of keratinolytics

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3. Fig. 2. Phylogenetic dendrogram based on the sequences of the internal transcribed spacers ITS1 and ITS2 of the rRNA genes. The phylogenetic tree was constructed using the maximum likelihood method in MEGA 10.0. Bootstrap analysis was performed using 1000 replicates. Values ​​greater than 70% are indicated.

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4. Fig. 3. Proteolytic activity of the studied micromycetes on media containing different nitrogen sources: C. pseudocladosporioides C66 – keratinolytic (a) and caseinolytic (b) activity; P. sizovae C11 – keratinolytic (c) and caseinolytic (d) activity; A. ochraceus ST2 – keratinolytic (e) and caseinolytic (f) activity; A. versicolor C51 – keratinolytic (g) and caseinolytic (h); K. terreum C106 – keratinolytic (i) and caseinolytic (j); T. inflatum ST1 – keratinolytic (k) and caseinolytic (l).

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5. Fig. 4. Dynamics of accumulation of extracellular proteases of T. inflatum ST1 culture during submerged cultivation on a medium without fish meal hydrolysate: 1 – keratinolysis; 2 – caseinolysis

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6. Fig. 5. Isoelectric focusing of extracellular proteins of the complex preparation T. inflatum ST1 in the pH range of ampholines 3.0–10.0: 1 – keratinolysis; 2 – caseinolysis; 3 – protein; 4 – pH

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7. Fig. 6. Electropherogram of keratinase T. inflatum ST1 (denaturing electrophoresis in PAGE).

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