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Creation of Chitinase Producer and Disruption of Micromycete Cell Wall with the Obtained Enzyme Preparation

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Abstract

A recombinant strain producing a complex of extracellular enzymes including chitinase from Myceliophtora thermophila was created based on the fungus Penicillium verruculosum. The activity of the enzyme preparations obtained from the cultural fluid of the producer strain was 0.55, 0.53, and 0.66 U/mg protein with chitin and chitosans with the molecular weight of 200 and 1000 kDa, respectively. The temperature optimum for the recombinant chitinase was 52-65°C; the pH optimum was 4.5-6.2, which corresponded to the published data for this class of the enzymes. The content of heterologous chitinase in the obtained enzyme preparations was 47% of total protein content in the cultural fluid. Enzyme preparations produced by the recombinant P. verruculosum XT403 strain and containing heterologous chitinase were able to degrade the mycelium of micromycetes, including phytopathogenic ones, and were very efficient in the bioconversion of microbiological industry waste.

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Abbreviations

CBHI:

cellobiohydrolase I

CF:

cultural fluid

CHT:

chitinase

DD:

degree of deacetylation

EG:

endoglucanase

EP:

enzyme preparation

XYL:

xylanase

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Acknowledgements

We used the equipment of the Industrial Biotechnologies Center for Collective Use of the Federal Research Center of Biotechnology, Russian Academy of Sciences.

Funding

This work was supported by the Ministry of Science and Higher Education (unique project identifier RFMEFI61620X0128).

Author information

Authors and Affiliations

  1. Lomonosov Moscow State University, Faculty of Chemistry, 119991, Moscow, Russia

    O. A. Sinitsyna & A. P. Sinitsyn

  2. Fundamentals of Biotechnology Federal Research Center, Russian Academy of Sciences, 119071, Moscow, Russia

    E. A. Rubtsova, I. G. Sinelnikov, D. O. Osipov, A. M. Rozhkova & A. P. Sinitsyn

  3. Pushchino Scientific Center for Biological Research, Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 142290, Pushchino, Moscow Region, Russia

    V. Yu. Matys, T. V. Bubnova & V. A. Nemashkalov

  4. Russian Research Institute of Food Biotechnology, 111033, Moscow, Russia

    A. S. Sereda

  5. All-Russian Research Institute of Phytopathology, 143050, Bolshye Vyazemy, Moscow Region, Russia

    L. A. Tcsherbakova

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  1. O. A. Sinitsyna
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Correspondence to O. A. Sinitsyna.

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Sinitsyna, O., Rubtsova, E., Sinelnikov, I. et al. Creation of Chitinase Producer and Disruption of Micromycete Cell Wall with the Obtained Enzyme Preparation. Biochemistry Moscow 85, 717–724 (2020). https://doi.org/10.1134/S0006297920060097

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