Comparative studies on trifluoroethanol (TFE) state of a thermophilic alpha-amylase and its mesophilic counterpart: limited proteolysis, conformational analysis, aggregation and reactivation of the enzymes

International Journal of Biological Macromolecules
S Mohsen AsghariHossein Naderi-Manesh

Abstract

Detailed circular dichroism (CD), scattering and quenching studies, 1-anilinonaphthalene-8-sulfonate (ANS) binding, irreversible thermoinactivation, activity measurements and proteolytic digestion of bacterial alpha-amylases have been carried out to elucidate the effect of trifluoroethanol (TFE) on the structure of these enzymes. Under high concentrations of TFE both of the alpha-amylases, a thermostable alpha-amylase from Bacillus licheniformis (BLA) and its mesophilic counterpart from Bacillus amyloliquefaciens (BAA), acquire partially folded state characterized by an enhanced content of the secondary structure (helix) and reduced tertiary structures. According to ANS binding studies, we suggest that the TFE states induced by TFE/water mixture are not the molten globule state in the alpha-amylase folding pathway. In addition, data shows significant reversible aggregation of both enzymes in TFE/water mixtures with concentration between 10 and 60% (v/v). However, reversibility is more in case of BAA. As expected, in the absence of TFE, the thermophilic enzyme compared to mesophilic enzyme, shows a greater resistance to digestion by thermolysin. With respect to fluorescence quenching by acrylamide and potassium iodide, the therm...Continue Reading

References

Dec 13, 1977·Biochemistry·M R Eftink, C A Ghiron
Dec 5, 1991·Journal of Molecular Biology·Y Goto, S Nishikiori
Apr 10, 1995·FEBS Letters·A FontanaP Polverino de Laureto
Nov 1, 1996·The Journal of Biological Chemistry·K RajaramanC M Rao
Mar 1, 1997·Trends in Biotechnology·A M Klibanov
Apr 1, 1997·Protein Science : a Publication of the Protein Society·P Polverino de LauretoA Fontana
Jan 1, 1997·Folding & Design·A FontanaM Zambonin
May 1, 1997·Journal of Protein Chemistry·T SivaramanC Yu
Mar 21, 1998·Proceedings of the National Academy of Sciences of the United States of America·V S Pande, D S Rokhsar
Apr 16, 1998·Biochimica Et Biophysica Acta·S J Hubbard
Jun 24, 1998·Proceedings of the National Academy of Sciences of the United States of America·P ZávodszkyG A Petsko
Mar 15, 2000·Proceedings of the National Academy of Sciences of the United States of America·G HernandezD M LeMaster
Mar 29, 2000·Proceedings of the National Academy of Sciences of the United States of America·R Jaenicke
Apr 4, 2000·Biotechnology and Applied Biochemistry·A PandeyR Mohan
Mar 22, 2001·Applied Biochemistry and Biotechnology·K Khajeh, M Nemat-Gorgani
Mar 27, 2001·Enzyme and Microbial Technology·K KhajehM Nemat-Gorgani
Apr 28, 2001·The Journal of Biological Chemistry·S D'AmicoG Feller
Jul 18, 2001·Applied Biochemistry and Biotechnology·K KhajehM Nemat-Gorgani
Jan 4, 2003·The Journal of Biological Chemistry·Salvino D'AmicoGeorges Feller
May 6, 2003·Archives of Biochemistry and Biophysics·Pawan GuptaM Saleemuddin

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Citations

Mar 7, 2006·Biophysical Chemistry·Maryam Monsef ShokriMehdi Sadeghi
Nov 30, 2005·Journal of Pharmaceutical Sciences·Ge JiangVincent J Sullivan

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