Vol 33(2017) N 6 p. 83-88; DOI 10.21519/0234-2758-2017-33-6-83-88
T.L. Gordeeva1*, L.N. Borshchevskaya1, A.N. Kalinina1, S.P. Sineoky1, S.P. Voronin2 and M.D. Kashirskaya2

Comparative Analysis in Plate Test of Expression Efficiency of Genes for Bacterial Phytases in Pichia pastoris yeast

1The State Research Institute for Genetics and Selection of Industrial Microorganisms of National Research Centr «Kurchatov Institute» (NRC «Kurchatov Institute» - GosNIIgenetika), 117545, Moscow Russia
2Bioamid, JSC, 410033, Saratov Russia

Received - 24.10.2017; Accepted - 13.11.2017


A comparative analysis of the efficiency of the Escherichia coli, Obessumbacterium proteus and Citrobacter freundii phytase gene expression in a Pichia pastoris methylotrophic yeast has been carried out. The level of the expression was assessed using a suggested for the first time simple model using a genetic construct based on the pPICά vector and a plate test with a modified diagnostic medium containing methanol instead of glucose and calcium phytate. The optimum pH value for the phytase reaction was provided by the addition of lactic acid. It was shown that the gene for the C. freundii phytase manifested the best expression among the studied heterological genes.

phytase, Pichia pastoris, plate test.

The work was financially supported by the Ministry of Education and Science of RF (Project Unique Identifier RFMEFI57917X0145) and used the help of the National Bioresource Center All-Russian Collection of Industrial Microorganisms (Moscow, Russia).