Effect of heat treatment on proper oligomeric structure formation of thermostable glutamate dehydrogenase from a hyperthermophilic archaeon

Biochemical and Biophysical Research Communications
R N Abd RahmanT Imanaka

Abstract

Natural glutamate dehydrogenase (Pk-GDH) was purified from hyperthermophilic archaeon Pyrococcus sp. KOD1 to homogeneity and its activity and structure were compared with those of recombinant enzyme, which was expressed in Escherichia coli. Determination of the molecular weight of these enzymes by SDS-PAGE and gel filtration revealed that the natural enzyme was purified only as a hexameric form, whereas the recombinant enzyme was purified as both monomeric and hexameric forms. Determination of the enzymatic activities indicated that only the enzyme in a hexameric form is active. Moreover, it is noted that the specific activity of the hexameric form of the recombinant enzyme is much lower than that of the natural enzyme and that circular dichroism spectra of these enzymes are distinctly different from each other. These results suggest that the structure of the hexameric form of the recombinant enzyme with low specific activity (Type I) is different from that of the natural enzyme with high specific activity (Type II). Upon heat treatment (80 degrees C, 15 min), the Type I structure was effectively converted to Type II structure and the specific activity of the enzyme was increased by 2.6-fold. Likewise, upon heat treatment (70 d...Continue Reading

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Citations

Apr 27, 2010·Protein Engineering, Design & Selection : PEDS·Peter Q Nguyen, Jonathan J Silberg
Nov 15, 2011·Proceedings of the Japan Academy. Series B, Physical and Biological Sciences·Tadayuki Imanaka
Oct 29, 2015·Extremophiles : Life Under Extreme Conditions·Ghazaleh GharibTadayuki Imanaka
Oct 20, 2005·Journal of Bioscience and Bioengineering·Naoshige IzumikawaMasahiro Takagi
May 30, 2006·Journal of Biotechnology·Takashi EndohTadayuki Imanaka
Jun 14, 2005·Biophysical Chemistry·Shingo NishikoriMasahiro Takagi
Oct 13, 2000·European Journal of Biochemistry·X F TangT Imanaka
Feb 20, 2007·Extremophiles : Life Under Extreme Conditions·Myoung-Hee LeeKwan-Hwa Park
Jul 11, 2012·Biotechnology Letters·Kosuke MinamihataNoriho Kamiya
Jul 12, 2002·Chemical Record : an Official Publication of the Chemical Society of Japan ... [et Al.]·Tadayuki Imanaka, Haruyuki Atomi
Nov 5, 1999·Biochemical and Biophysical Research Communications·T YamamotoT Imanaka
Aug 15, 1998·Biochemical and Biophysical Research Communications·R N RahmanT Imanaka

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