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

O. A. Sinitsyna1,a*, E. A. Rubtsova2, I. G. Sinelnikov2, D. O. Osipov2, A. M. Rozhkova2, V. Yu. Matys3, T. V. Bubnova3, V. A. Nemashkalov3, A. S. Sereda4, L. A. Tcsherbakova5, and A. P. Sinitsyn1,2

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

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

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

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

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

* To whom correspondence should be addressed.

Received April 16, 2020; Revised April 18, 2020; Accepted April 18, 2020
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.
KEY WORDS: chitinase, chitin, chitosan, Penicillium verruculosum, phytopathogenic fungi, waste bioconversion

DOI: 10.1134/S0006297920060097