Improvement of protein stability and enzyme recovery under stress conditions by using a small HSP (tpv-HSP 14.3) from Thermoplasma volcanium

Date

2012-11-01

Author

Kocabıyık, Semra

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In this study we cloned and expressed a small heat shock protein, tpv-HSP 14.3, from thermoacidophilic archaeon Thermoplasma volcanium. This novel recombinant small heat shock protein was purified to homogeneity and produced a protein band of 14.3 kDa on SDS-polyacrylamide gel. Transmission electron microscopy images of the negatively stained tpv-EISP 14.3 samples showed spherical particles of 13 nm diameter. E. coli cells over expressing tpv-HSP 14.3 endowed the cells with some degree of thermotolerance. After exposure to 52 degrees C for 120 min, survivability of the E. coli cells expressing tpv-HSP 14.3 was approximately 2.5-fold higher than the control cells. As a molecular chaperone tpv-HSP 14.3 enhanced the thermal stabilization of substrate proteins, pig heart citrate synthase and bovine L-glutamic dehdyrogenase, considerably. The highest protection effect of tpv-HSP 14.3 was observed at 47 degrees C for pig heart citrate synthase; the remaining activity was 5-fold higher than that of the sample without tpv-HSP 14.3. The tpv-sHSP 14.3 prevented inactivation of bovine L-glutamic dehdyrogenase the most effectively at 53 degrees C; the residual activity was approximately 2-fold higher than that of the sample heated without tpv-HSP 14.3. However, refolding activity of the tpv-HSP 14.3 was relatively weak for the chemically denatured substrate proteins.

Subject Keywords

Thermoplasma volcanium, Small heat-shock protein (sHSP), Molecular chaperone, Heat-shock response

URI

https://hdl.handle.net/11511/38474

Journal

PROCESS BIOCHEMISTRY

DOI

https://doi.org/10.1016/j.procbio.2011.11.014

Collections

Department of Biology, Article

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Citation Formats

S. Kocabıyık, “Improvement of protein stability and enzyme recovery under stress conditions by using a small HSP (tpv-HSP 14.3) from Thermoplasma volcanium,” PROCESS BIOCHEMISTRY, pp. 1676–1683, 2012, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/38474.