Reference : Temperature adaptation of proteins: Engineering mesophilic-like activity and stabilit...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
Temperature adaptation of proteins: Engineering mesophilic-like activity and stability in a cold-adapted alpha-amylase
D'Amico, Salvino [Université de Liège - ULg > > GIGA-Research >]
Gerday, Charles mailto [Université de Liège - ULg > Services généraux (Faculté des sciences) > Relations académiques et scientifiques (Sciences) >]
Feller, Georges mailto [Université de Liège - ULg > Département des sciences de la vie > Labo de biochimie >]
Journal of Molecular Biology
Academic Press Ltd Elsevier Science Ltd
Yes (verified by ORBi)
[en] psychrophile ; cold adaptation ; alpha-amylase ; thermal stability ; microcalorimetry
[en] Two multiple mutants of a psychrophilic alpha-amylase were produced, bearing five mutations (each introducing additional weak interactions found in pig pancreatic (alpha-amylase) with or without an extra disulfide bond specific to warm-blooded animals. Both multiple mutants display large modifications of stability and activity arising from synergic effects in comparison with single mutations. Newly introduced weak interactions and the disulfide bond confer mesophilic-like stability parameters, as shown by increases in the melting point t(m), in the calorimetric enthalpy DeltaH(cal) and in protection against heat inactivation, as well as by decreases in cooperativity and reversibility of unfolding. In addition, both kinetic and thermodynamic activation parameters of the catalyzed reaction are shifted close to the values of the porcine enzyme. This study confirms the central role of weak interactions in regulating the balance between stability and activity of an enzyme in order to adapt to the environmental temperature. (C) 2003 Elsevier Ltd. All rights reserved.

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