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Vol 34(2018) N 4 p. 18-25; DOI 10.21519/0234-2758-2018-34-4-18-25
T.L. Gordeeva1*, L.N. Borshchevskaya1, A.N. Kalinina1, S.P. Sineoky1, S.P. Voronin2, M.D. Kashirskaya2

Expression and Characteristics of Phytases from Obesumbacterium proteus in Pichia pastoris Yeast

1State Research Institute for Genetics and Selection of Industrial Microorganisms of National Research Center «Kurchatov Institute» (NRC «Kurchatov Institute» - GOSNIIGENETIKA), 117545, Moscow Russia
2Joint Venture Bioamide, 410033, Saratov Russia

*tatiana.gordeeva@mail.ru
Received - 24.05.2018; Accepted - 06.07.2018

References

In order to increase the level of expression of recombinant phytases in the Pichia pastoris yeast, a synthetic sequence of the phyOp-mod gene that encodes phytase from Obesumbacterium proteus has been developed and optimized by codon composition. The GC content was adjusted to 48.33, and the AT-rich segments were eliminated. The main characteristics for the purified enzyme were established. The optimal pH and temperature values for the phytase РhyOp functioning proved to be equal to 4.0 and 50 °C, respectively. The specific activity, Michaelis constant (Km) and maximal rate of reaction (Vmax) with sodium phytate as a substrate was 1452 U/mg, 0.31 mM and 823 U/mg, respectively. It was shown that the optimization of the codon composition increased in the expression of the target product. The recombinant phytase designated as PhyOp demonstrated great potential for application as a feed additive due to its relatively high specific activity and good kinetic characteristics.

phytase from Obesumbacterium proteus, Pichia pastoris, codon optimization.

The work was financially supported by the Ministry of Education and Science of RF (Unique Identifier of the Project RFMEFI57917X0145) using Multipurpose Scientific Installation of National Bioresource Center All-Russian Collection of Industrial Microorganisms, NRC «Kurchatov Institute» - GOSNIIGENETIKA.



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