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New Recombinant Phytase from Kosakoniasacchari: Characteristics and Biotechnological Potential

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Abstract

A DNA sequence from Kosakonia sacchari that, according to automated computer analysis, is believed to correspond to the gene for histidine-acid phytase has been selected from the GenBank database. The sequence was optimized for codon composition, synthesized, cloned, and expressed in Pichia pastoris. The main characteristics of the purified recombinant enzyme were determined. It was established that a pH of 4.5 and a temperature of 50°C are optimal for phytase functioning. The specific activity, Michaelis constant (Km), and maximum reaction rate (Vmax) with phytate as a substrate were 1470 U/mg, 193 μM and 2167 μmol/(min mg), respectively. It was shown that the enzyme was characterized by a wide range of working pH levels. Therefore, due to its properties, this new recombinant phytase can be considered a high-potential enzyme for agrobiotechnology.

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ACKNOWLEDGEMENTS

The work was carried out at the Unique Scientific Facility of the All-Russia Collection of Industrial Microorganisms National Bioresource Center, Kurchatov Institute NRC, GOSNIIGENETIKA.

Funding

The work was financially supported by the Ministry of Education and Science of Russian Federation (Unique Project Identifier RFMEFI57917X0145).

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Authors and Affiliations

  1. State Research Institute for Genetics and Selection of Industrial Microorganisms, Kurchatov Institute National Research Center (Kurchatov Institute NRC, GOSNIIGENETIKA), 117545, Moscow, Russia

    T. L. Gordeeva, L. N. Borshchevskaya, A. N. Kalinina, N. V. Bulushova & S. P. Syneoky

  2. BIOAMIDE Closed Joint Stock Co., 410033, Saratov, Russia

    S. P. Voronin & M. D. Kashirskaya

Authors
  1. T. L. Gordeeva
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  2. L. N. Borshchevskaya
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  3. A. N. Kalinina
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  4. N. V. Bulushova
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  5. S. P. Syneoky
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  6. S. P. Voronin
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  7. M. D. Kashirskaya
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Corresponding author

Correspondence to T. L. Gordeeva.

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This article does not contain any studies involving animals or humans performed by any of the authors.

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Translated by I. Gordon

Abbreviations: aa—amino-acid residue(s); CAI—codon adaptation index; CL—culture liquid; PAGE—polyacrylamide gel electrophoresis; PCR—polymerase chain reaction; SDS—sodium dodecyl(lauryl)sulfate.

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Gordeeva, T.L., Borshchevskaya, L.N., Kalinina, A.N. et al. New Recombinant Phytase from Kosakoniasacchari: Characteristics and Biotechnological Potential. Appl Biochem Microbiol 56, 779–786 (2020). https://doi.org/10.1134/S0003683820070042

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