Kinetic study of the thermal denaturation of a hyperthermostable extracellular α-amylase from Pyrococcus furiosus

被引：23

作者：

Brown, I. ^{[1
]}

Dafforn, T. R. ^{[2
]}

Fryer, P. J. ^{[1
]}

Cox, P. W. ^{[1
]}

机构：

[1] Univ Birmingham, Sch Chem Engn, Birmingham B15 2TT, W Midlands, England

[2] Univ Birmingham, Sch Biosci, Birmingham B15 2TT, W Midlands, England

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS | 2013年 / 1834卷 / 12期

基金：

英国工程与自然科学研究理事会;

关键词：

Pyrococcus furiosus; alpha-Amylase; Sterilisation; TTI; Denaturation; AMYLOLYTIC ENZYMES; GLOBULAR-PROTEINS; STABILITY; STABILIZATION; DEACTIVATION; PURIFICATION; MECHANISM; ARCHAEON;

D O I：

10.1016/j.bbapap.2013.09.008

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Hyperthermophilic enzymes are of industrial importance and interest, especially due to their denaturation kinetics at commercial sterilisation temperatures inside safety indicating time-temperature integrators (TTIs). The thermal stability and irreversible thermal inactivation of native extracellular Pyrococcus furiosus a-amylase were investigated using differential scanning calorimetiy, circular dichroism and Fourier transform infrared spectroscopy. Denaturation of the amylase was irreversible above a T-m of approximately 106 degrees C and could be described by a one-step irreversible model. The activation energy at 121 degrees C was found to be 316 kJ/mol. Using CD and FT-IR spectroscopy it was shown that folding and stability greatly increase with temperature. Under an isothermal holding temperature of 121 degrees C, the structure of the PFA changes during denaturation from an alpha-helical structure, through a beta-sheet structure to an aggregated protein. Such data reinforces the use of P. furiosus alpha-amylase as a labile species in TTIs. (C) 2013 Elsevier B.V. All rights reserved.

引用

页码：2600 / 2605

页数：6

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