Temperature adaptation of proteins: engineering mesophilic-like activity and stability in a cold-adapted alpha-amylase.
Abstract
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. Study holds ProTherm entries: 16782, 16783, 16784, 16785, 16786 Extra Details: psychrophile; cold adaptation; alpha-amylase; thermal stability; microcalorimetry
Submission Details
ID: B26v8nVV
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:48 p.m.
Version: 1
Publication Details
D'Amico S;Gerday C;Feller G,J. Mol. Biol. (2003) Temperature adaptation of proteins: engineering mesophilic-like activity and stability in a cold-adapted alpha-amylase. PMID:14499602Additional Information
Study Summary
Structure view and single mutant data analysis
Study data
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Data Distribution
Studies with similar sequences (approximate matches)