Preparation and characterization of a new mutant homolog of chemotaxis protein CheY from anaerobic hyperthermophilic microorganism Thermotoga naphthophila

   
Grishin D.V.1 , Gladilina Ju.A.1, Zhdanov D.D.2, Pokrovskaya M.V.1, Toropygin I.Yu.1, Aleksandrova S.S.1, Pokrovskiy V.S.2, Sokolov N.N.1

1. Institute of Biomedical Chemistry, Moscow, Russia
2. Institute of Biomedical Chemistry, Moscow, Russia; Peoples` Friendship University of Russia, Moscow, Russia
Section: Experimental Study
DOI: 10.18097/PBMC20196501041      PubMed Id: 30816096
Year: 2019  Volume: 65  Issue: 1  Pages: 41-50
Using genetic engineering methods the expression vectors structures have been designed to produce recombinant proteins TnaCheY and Tna CheY-mut, the homologues of the chemotaxis protein CheY from the hyperthermophilic organism Thermotoga naphthophila in Escherichia coli BL21(DE3) cells. The cultivation conditions of transformed strains were optimized. The influence of episomal expression of the heterologous chemotaxis protein CheY on growth kinetics parameters of the culture of mesophilic bacteria E. coli was studied. The optimal purification flowchart of the obtained proteins using thermolysis is proposed. Using the E. coli BL21(DE3) laboratory strain as an example, the possibility of employment the episomal expression of such proteins to control the cultivation and production time of pharmaceutically and industrially valuable metabolites due to the impact on some stages of the bacterial chemotaxis is experimentally proved.
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Keywords: recombinant protein, thermal stability, chemotaxis protein CheY, strain-producer, expression, biotechnology
Citation:

Grishin, D. V., Gladilina, Ju. A., Zhdanov, D. D., Pokrovskaya, M. V., Toropygin, I. Yu., Aleksandrova, S. S., Pokrovskiy, V. S., Sokolov, N. N. (2019). Preparation and characterization of a new mutant homolog of chemotaxis protein CheY from anaerobic hyperthermophilic microorganism Thermotoga naphthophila. Biomeditsinskaya khimiya, 65(1), 41-50.
This paper is also available as the English translation: 10.1134/S1990750819030041
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