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:14499602
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