Cellulase activity of mycelial fungi collection strains study

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Abstract

Relevance. Cellulases occupy the third place in the world among industrially produced enzymes. This is largely due to the fact that cellulose is the main component of plant material, and the lignin-cellulose part of biomass accumulates in huge quantities in the form of waste from agriculture, woodworking and other industries. To convert this material, it is necessary to perform its enzymatic cleavage to glucose and cellobiose using various cellulases.

The purpose of this work was to study the cellulose activity of collection strains of mycelial fungi for the selection of promising strains-producers of cellulases

Material and methods. The objects of the study were 13 strains of 12 species of micromycetes from the microorganisms biocollection of the VILAR, belonging to the genera Aspergillus, Monilia, Penicillium. The work used surface and deep cultivation of fungi on media with partial replacement of sucrose with cellulose. The cellulolytic activity of microorganisms was assessed by the growth rate of colonies. In addition, at the last stage of cultivation, the surface of the agar was stained with Lugol solution, the diameter of the lysis zones was measured and lysis indices were calculated.

Mushroom cultivation in deep conditions was carried out in flasks on a shaker. The seed material was a suspension of spores of seven-day deuteromycete cultures. In the culture fluid filtrates, the total cellulase activity was evaluated by determining reducing sugars, as well as the concentration of sucrose.

Statistical processing of the results, regression and correlation analysis were carried out on a personal computer using the Microsoft Office Excel 2010 statistical software package.

Results. During surface cultivation on a modified medium with cellulose, fungi formed colonies and well-defined lysis zones, which indicated the synthesis and secretion of cellulolytic enzymes. Differences in radial growth rates and lysis indices were found in individual species and strains of micromycetes. Using regression and correlation analysis, six strains were selected for deep cultivation. The presence of hydrolytic activity with respect to microcrystalline cellulose in the culture fluid of fungi during cultivation on a medium with partial replacement of sucrose with cellulose is shown.

Conclusions. The synthesis of cellulases was found by the studied fungal cultures during surface and deep cultivation. A comprehensive analysis of the data obtained makes it possible to select the most promising cellulase-producing strains.

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About the authors

Z. K. Nikitina

All-Russian Scientific Institute of Medicinal and Aromatic Plants

Author for correspondence.
Email: nikitinaz@yandex.ru

Dr.Sc. (Biol.), Professor

Russian Federation, Moscow

I. K. Gordonova

All-Russian Scientific Institute of Medicinal and Aromatic Plants

Email: gordonova777@yandex.ru

Ph.D. (Biol.), Leading Research Scientist

Russian Federation, Moscow

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Supplementary files

Supplementary Files
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1. JATS XML
2. Figure 1. Growth of micromycetes on media with cellulose: upper row: P. malinovobranova F 3 (4th, 5th, and 6th days), lower row, P. bre-vicompactum F 37 (4th, 5th, and 6th day)

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3. Figure 2. Changes in the radial growth rate of fungi during cultivation on cellulose media (top shows cultivation time in days)

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4. Figure 3. Maximum rates of radial growth of micromycetes during cultivation on cellulose media

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5. Figure 4. Colonies and zones of lysis of fungi during cultivation on media with cellulose. From left to right: P. purpurescens F 18 without color, P. purpurescens F 18 with color, P. brevicompactum F 37, A. terries F 59

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6. Figure 5. Indices of fungal lysis during growth on media containing cellulose

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7. Figure 6. Graph and linear regression equation showing the relationship between maximum growth rate and micromycete lysis index (R2 - coefficient of determination)

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