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

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@article{fd545392675144e49f126f9c2106f919,
title = "Kinetic study of the thermal denaturation of a hyperthermostable extracellular α-amylase from Pyrococcus furiosus",
abstract = "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 α-amylase were investigated using differential scanning calorimetry, circular dichroism and Fourier transform infrared spectroscopy. Denaturation of the amylase was irreversible above a Tm of approximately 106 °C and could be described by a one-step irreversible model. The activation energy at 121 °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 °C, the structure of the PFA changes during denaturation from an α-helical structure, through a β-sheet structure to an aggregated protein. Such data reinforces the use of P. furiosus α-amylase as a labile species in TTIs.",
keywords = "Pyrococcus furiosus, α-Amylase, Sterilisation, TTI, Denaturation",
author = "Ian Brown and Timothy Dafforn and Peter Fryer and Philip Cox",
year = "2013",
month = dec,
doi = "10.1016/j.bbapap.2013.09.008",
language = "English",
volume = "1834",
pages = "2600--2605",
journal = "Biochimica et Biophysica Acta. Proteins and Proteomics",
issn = "1570-9639",
publisher = "Elsevier",
number = "12",

}

RIS

TY - JOUR

T1 - Kinetic study of the thermal denaturation of a hyperthermostable extracellular α-amylase from Pyrococcus furiosus

AU - Brown, Ian

AU - Dafforn, Timothy

AU - Fryer, Peter

AU - Cox, Philip

PY - 2013/12

Y1 - 2013/12

N2 - 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 α-amylase were investigated using differential scanning calorimetry, circular dichroism and Fourier transform infrared spectroscopy. Denaturation of the amylase was irreversible above a Tm of approximately 106 °C and could be described by a one-step irreversible model. The activation energy at 121 °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 °C, the structure of the PFA changes during denaturation from an α-helical structure, through a β-sheet structure to an aggregated protein. Such data reinforces the use of P. furiosus α-amylase as a labile species in TTIs.

AB - 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 α-amylase were investigated using differential scanning calorimetry, circular dichroism and Fourier transform infrared spectroscopy. Denaturation of the amylase was irreversible above a Tm of approximately 106 °C and could be described by a one-step irreversible model. The activation energy at 121 °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 °C, the structure of the PFA changes during denaturation from an α-helical structure, through a β-sheet structure to an aggregated protein. Such data reinforces the use of P. furiosus α-amylase as a labile species in TTIs.

KW - Pyrococcus furiosus

KW - α-Amylase

KW - Sterilisation

KW - TTI

KW - Denaturation

U2 - 10.1016/j.bbapap.2013.09.008

DO - 10.1016/j.bbapap.2013.09.008

M3 - Article

C2 - 24063888

VL - 1834

SP - 2600

EP - 2605

JO - Biochimica et Biophysica Acta. Proteins and Proteomics

JF - Biochimica et Biophysica Acta. Proteins and Proteomics

SN - 1570-9639

IS - 12

ER -