Assessment of the efficiency of recombinant strains-producers of Bacillus pumilus 3-19 metalloendopeptidase

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Abstract

Abstract. Minor zinc-dependent metalloendopeptidase secreted by the soil strain Bacillus pumilus 3-19 occupies a unique intermediate classification position between two families of the metzincin clan: adamalizins and astacins. To be able to study the functional role of metalloendopeptidase in more detail, it is necessary to obtain a pure protein preparation in sufficient quantity. Since the enzyme secretion by the B. pumilus 3-19 strain is extremely low, recombinant strains producing metalloendopeptidase MprBp were constructed based on protease-free strains of Bacillus subtilis, as well as methylotrophic yeast Pichia pastoris. The obtained strains were evaluated for the efficiency of metalloendopeptidase production. The B. subtilis strain BG2036+mprBp had the highest production of the target protein.

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About the authors

N. L. Rudakova

Kazan (Volga Region) Federal University

Author for correspondence.
Email: natalialrudakova@mail.ru

Institute of Fundamental Medicine and Biology

Russian Federation, 420008, Kazan

D. I. Khasanov

Kazan (Volga Region) Federal University

Email: natalialrudakova@mail.ru

Institute of Fundamental Medicine and Biology

Russian Federation, 420008, Kazan

M. R. Sharipova

Kazan (Volga Region) Federal University

Email: natalialrudakova@mail.ru

Institute of Fundamental Medicine and Biology

Russian Federation, 420008, Kazan

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2. Figure. Dynamics of growth and accumulation of proteolytic activity by the B. pumilus 3-19 strain. The dispersion value for the culture growth index (OD600) does not exceed 8%, for the level of proteolytic activity the dispersion does not exceed 10%. Statistically significant differences in the level of MprBp activity from the total level of proteolytic activity are noted * (P < 0.05).

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