Denaturing Effects of Urea and Guanidine Hydrochloride on Hyperthermophilic Esterase from Aeropyrum pernix K1

Chemical Research in Chinese Universities ›› 2006, Vol. 22 ›› Issue (2): 168-172.

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Denaturing Effects of Urea and Guanidine Hydrochloride on Hyperthermophilic Esterase from Aeropyrum pernix K1

GAO Ren-jun¹, XIE Gui-qiu^1,2, ZHOU Jun¹, FENG Yan¹, CAO Shu-gui¹

1. Key Laboratory for Molecular Enzymology and Engineering, the Ministry of Education;Jilin University, Changchun 130021, P.R.China;

2. School of Pharmacy, Jilin University, Changchun 130021, P.R.China

Received:2005-12-20 Online:2006-04-24 Published:2011-08-06
Supported by:
Supported by the National Natural Science Foundation of China(Nos.30400081,20272017 and 30570405) and the Key Technologies Research and Development Programmer (No.2001 BA708 B03-06).

Abstract

Abstract: The changes in the activity and the conformation of the hyperthermophilic esterase derived from aerobic thermophilic Aeropyrumpernix K1 (APE1547) were studied during denaturation by guanidine hydrochloride (GdnHCl)and urea.The denaturation course of APE1547 was followed by the steady-state and time resolved fluorescence methods.An increase in the denaturant concentration in the denatured system can significantly enhance the inactivation and unfolding of APE1547.The enzyme can be completely inactivated with a urea concentration of 2.7 mol/L or a GdnHCl concentration of 7.5 mol/L.The fluorescence emission maximum of the enzyme protein red shifts in magnitude to a maximum value(355 nm) when the concentration of GdnHCl is 5.1 mol/L.The experimental results indicate that APE1547 has a high resistance to urea.Unfolding of APE1547 in GdnHCl(4.2-6.0 mol/L) was shown to be an irreversible process.The present results indicate that the ion pairs in this protein may be a key factor for the stability of this esterase.

Key words: Stability, Hyperthermophilic esterase, Urea, Guanidine hydrochloride

GAO Ren-jun, XIE Gui-qiu, ZHOU Jun, FENG Yan, CAO Shu-gui. Denaturing Effects of Urea and Guanidine Hydrochloride on Hyperthermophilic Esterase from Aeropyrum pernix K1[J]. Chemical Research in Chinese Universities, 2006, 22(2): 168-172.

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Denaturing Effects of Urea and Guanidine Hydrochloride on Hyperthermophilic Esterase from Aeropyrum pernix K1

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