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Kinetic study of the thermal denaturation of a hyperthermostable extracellular α-amylase from Pyrococcus furiosus.
Biochim Biophys Acta. 2013 Dec; 1834(12):2600-5.BB

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 316kJ/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.

Authors+Show Affiliations

School of Chemical Engineering, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24063888

Citation

Brown, I, et al. "Kinetic Study of the Thermal Denaturation of a Hyperthermostable Extracellular Α-amylase From Pyrococcus Furiosus." Biochimica Et Biophysica Acta, vol. 1834, no. 12, 2013, pp. 2600-5.
Brown I, Dafforn TR, Fryer PJ, et al. Kinetic study of the thermal denaturation of a hyperthermostable extracellular α-amylase from Pyrococcus furiosus. Biochim Biophys Acta. 2013;1834(12):2600-5.
Brown, I., Dafforn, T. R., Fryer, P. J., & Cox, P. W. (2013). Kinetic study of the thermal denaturation of a hyperthermostable extracellular α-amylase from Pyrococcus furiosus. Biochimica Et Biophysica Acta, 1834(12), 2600-5. https://doi.org/10.1016/j.bbapap.2013.09.008
Brown I, et al. Kinetic Study of the Thermal Denaturation of a Hyperthermostable Extracellular Α-amylase From Pyrococcus Furiosus. Biochim Biophys Acta. 2013;1834(12):2600-5. PubMed PMID: 24063888.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Kinetic study of the thermal denaturation of a hyperthermostable extracellular α-amylase from Pyrococcus furiosus. AU - Brown,I, AU - Dafforn,T R, AU - Fryer,P J, AU - Cox,P W, Y1 - 2013/09/21/ PY - 2013/07/23/received PY - 2013/09/13/revised PY - 2013/09/14/accepted PY - 2013/9/26/entrez PY - 2013/9/26/pubmed PY - 2014/2/22/medline KW - Denaturation KW - Pyrococcus furiosus KW - Sterilisation KW - TTI KW - α-Amylase SP - 2600 EP - 5 JF - Biochimica et biophysica acta JO - Biochim Biophys Acta VL - 1834 IS - 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 316kJ/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. SN - 0006-3002 UR - https://www.unboundmedicine.com/medline/citation/24063888/Kinetic_study_of_the_thermal_denaturation_of_a_hyperthermostable_extracellular_α_amylase_from_Pyrococcus_furiosus_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1570-9639(13)00342-7 DB - PRIME DP - Unbound Medicine ER -