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Enhancement of Activity of Carbohydrases with Endo-depolymerase Action in Trichoderma reesei Using Mutagenesis

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Abstract—

In order to obtain feed enzyme preparations with increased activity of endo-carbohydrases of depolymerase action, mutagenesis of the Trichoderma reesei strain was carried out using ultraviolet and ionizing irradiation. Application of fractionated UV-irradiation made it possible to increase the endoglucanase activity by 20% and xylanase activity by 80% compared to the original strain. The greatest effect was achieved after multistage gamma-irradiation mutagenesis with a cobalt source. When the mutant T. reesei-Co-44 was cultured in shaking flasks, its endoglucanase activity was increased 5-fold, while xylanase activity increased more than 8-fold compared to the original strain. High activity of the target enzymes in the mutant strains was confirmed by the results of fed-batch cultivation in laboratory reactors with lactose feeding as a carbon source and an inducer of cellulases and xylanases biosynthesis. The maximum activity of endoglucanase and xylanase was achieved by culturing strain T. reesei-Co-44 obtained as a result of gamma-irradiation mutagenesis and amounted to 1324.2 ± 70.1 and 3394 ± 164 U/mL, respectively. This activity level makes it possible to produce competitive enzyme preparations of carbohydrases for fodder production.

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ACKNOWLEDGMENTS

This work was financially supported by a grant of the Basic Research Program of the Russian Academies of Sciences for 2013–2020 (project no. 0529-2016-0045); experiments were performed using the facilities of the Shared-Access Center “Industrial Biotechnologies” of the Research Center of Biotechnology, Russian Academy of Sciences.

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Authors and Affiliations

  1. All-Russian Research Institute of Food Biotechnology, a Branch of FRC of Food, Biotechnology, and Food Safety, Moscow, Russia

    E. V. Kostyleva, N. V. Tsurikova, A. S. Sereda, I. A. Velikoretskaya & T. N. Veselkina

  2. Kurchatov Institute, Moscow, Russia

    N. S. Lobanov

  3. Research Center of Biotechnology, Russian Academy of Sciences, Moscow, Russia

    I. A. Shashkov & A. P. Sinitsyn

  4. Chemical Faculty, Moscow State University, Moscow, Russia

    A. P. Sinitsyn

Authors
  1. E. V. Kostyleva
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  2. N. V. Tsurikova
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  3. A. S. Sereda
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  4. I. A. Velikoretskaya
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  5. T. N. Veselkina
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  6. N. S. Lobanov
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  7. I. A. Shashkov
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  8. A. P. Sinitsyn
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Correspondence to E. V. Kostyleva.

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Translated by D. Timchenko

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Kostyleva, E.V., Tsurikova, N.V., Sereda, A. et al. Enhancement of Activity of Carbohydrases with Endo-depolymerase Action in Trichoderma reesei Using Mutagenesis. Microbiology 87, 652–661 (2018). https://doi.org/10.1134/S0026261718050120

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