[Construction and characterization of a recombinant mutant homolog of the CheW protein from Thermotoga petrophila RKU-1]

Biomed Khim. 2018 Jan;64(1):53-60. doi: 10.18097/PBMC20186401053.
[Article in Russian]

Abstract

In the work a recombinant chemotaxis protein CheW from Thermotoga petrophila RKU-1 (TpeCheW) and its mutant homolog (TpeCheW-mut) were created. It was shown that, despite the low homology with CheW prototypes from intestinal bacteria, these proteins didn't cause metabolic overload and were well expressed by cells of E. coli laboratory strains. We have discovered a broad spectrum of industrial valuable properties of the TpeCheW-mut protein such as stability in a wide range of temperatures and pH, high expression level, solubility and possibility of the application of a simple low-stage purification methodology with the use of preliminary heat treatment. Possible directions of the scientific and industrial application of this protein were claimed.

V rabote polucheny rekombinantnyĭ khemotaksisnyĭ belok CheW Thermotoga petrophila RKU-1 (TpeCheW) i ego mutantnyĭ gomolog (TpeCheW-mut). Pokazano, chto, nesmotria na nevysokuiu gomologiiu s CheW bakteriĭ gruppy kishechnoĭ palochki, dannye belki ne vyzyvaiut metabolicheskoĭ peregruzki i khorosho ékspressiruiutsia kletkami laboratornykh shtammov E. coli. Vyiavlen shirokiĭ spektr vazhnykh dlia vydeleniia belka TpeCheW-mut svoĭstv i parametrov, takikh kak stabil'nost' v shirokom diapazone temperatur i znacheniĭ rN, vysokiĭ uroven' ékspressii, rastvorimost', a takzhe vozmozhnost' primeneniia prostykh malostadiĭnykh skhem ochistki, vkliuchaia termoobrabotku. Sformulirovany i obosnovany vozmozhnye napravleniia ispol'zovaniia dannogo belka v praktike nauchnykh i prikladnykh issledovaniĭ.

Keywords: biotechnology; hyperthermophilic microorganism; protein overexpression; recombinant thermostable protein CheW.

MeSH terms

  • Bacterial Proteins
  • Escherichia coli
  • Escherichia coli Proteins
  • Gram-Negative Anaerobic Straight, Curved, and Helical Rods*
  • Recombinant Proteins

Substances

  • Bacterial Proteins
  • CheW protein, E coli
  • Escherichia coli Proteins
  • Recombinant Proteins