Preparation and Characterization of a New Mutant Homolog of the Chemotaxis Protein CheY from the Anaerobic Hyperthermophilic Microorganism Thermotoga Naphthophila
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Using genetic engineering methods we have developed expression vectors for synthesis of recombinant proteins TnaCheY and TnaCheY-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 cell 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 investigated. The optimal purification flowchart of the obtained proteins using thermolysis has been proposed. Based on the data obtained, we discuss potential areas of application of recombinant variants of the CheY thermostable chemotactic protein. Using the E. coli BL21(DE3) laboratory strain as an example, the possibility of employment of 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 has been experimentally proven.
Keywords:recombinant protein thermal stability chemotaxis protein CheY strain-producer expression biotechnology
The study was financially supported by the Russian Science Foundation (project no. 16-16-04086).
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This article does not contain any research involving humans or using animals as objects.
- 3.Park, S.Y., Chao, X., Gonzalez-Bonet, G., Beel, B.D., Bilwes, A.M., and Crane, B.R., Mol. Cell, 2004, vol. 16, pp. 563–574.Google Scholar
- 12.Grishin, D.V., Podobed, O.V., Gladilina, Yu.A., Pokrovskaya, M.V., Aleksandrova, S.S., Pokrovskyi, V.S., and Sokolov, N.N., Vopr. Pitaniya, 2017, vol. 86, pp. 19–31.Google Scholar
- 13.Drury, L., Methods Mol. Biol., 1996, vol. 58, pp. 249–256.Google Scholar
- 15.Yadav, P., Yadav, A., Garg, V., Datta, T.K., Goswami, S.L., and De, S., Indian J. Exp. Biol., 2011, vol. 49, pp. 558–560.Google Scholar
- 23.Costa, S., Almeida, A., Castro, A., and Domingues, L., Front. Microbiol, 2014, vol. 19, 63.Google Scholar