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Cloning, Prokaryotic Expression, and Functional Characterization of a Novel 70-Kda Heat Shock Protein (Dnak) From Bacillus Persicus Publisher



Lotfi M1 ; Ghafouri H1, 2 ; Sarikhan S3 ; Shahangian SS1 ; Darvishi R4
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Authors Affiliations
  1. 1. Department of Biology, University of Guilan, Rasht, Iran
  2. 2. Department of Marine Sciences, The Caspian Sea Basin Research Center, University of Guilan, Rasht, Iran
  3. 3. Molecular Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran
  4. 4. Department of Medicine, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

Source: Process Biochemistry Published:2021


Abstract

The dnaK gene of Bacillus persicus strain B48(T) was cloned, sequenced, and functionally characterized in the present study. The gene was 1836 bp in length, encoding a polypeptide of 611 amino acid residues. The 3D structure of protein predicted by I-TASSER represented the similarity of the overall structures of DnaK from B. persicus strain B48(T) and human protein Hsp70 (BiP) with a homology of 89 %. Based on the results, refolding heat-denatured carbonic anhydrase increased significantly up to 80 % in the presence of the purified recombinant DnaK. In addition, salt resistance experiments demonstrated a 2.7-fold increase in the survival of recombinant E. coli BL21–DnaK in the presence of 0.4 M NaCl for 60 h compared to that of the control cells. Further, Cd2+ failed to affect DnaK refolding function, while Hg2+ ion reflected a biphasic effect (inhibiting and stimulating at lower and higher than 100 nM concentrations, respectively). Finally, DnaK from B. persicus can potentially be used for improving the functional properties of enzymes and proteins through increasing folding activity and enhancing stress tolerance. © 2020 Elsevier Ltd
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