The thermal stability of the Aes acetyl esterase from Escherichia coli has been investigated by means of differential scanning calorimetry and circular dichroism measurements. The calorimetric curves show a denaturation temperature of 68 °C for Aes and 61 °C for the single point mutant V20D-Aes. The same values are obtained from CD denaturation curves of the two proteins recorded in both the far-UV and near-UV regions. Even if the denaturation process is irreversible and characterized by a single calorimetric peak and a single inflection point in both far- and near-UV CD curves, the overall data indicate that the process is more complex than a two-state transition. This is in line with the presence of two structural domains in the 3D model of Aes, according to homology modelling. A comparison of the thermal stability of Aes with those of the homologous thermophilic EST2 and hyperthermophilic AFEST suggests that the optimization of charge–charge interactions should not be so effective in the case of the mesophilic enzyme.

Temperature-induced denaturation of Aes acetyl-esterase from Escherichia coli / DEL VECCHIO, POMPEA GIUSEPPINA GRAZIA; G., Graziano; Barone, Guido; L., Mandrich; Rossi, Mose'; G., Manco. - In: THERMOCHIMICA ACTA. - ISSN 0040-6031. - STAMPA. - 441:2(2006), pp. 144-149. [10.1016/j.tca.2005.12.006]

Temperature-induced denaturation of Aes acetyl-esterase from Escherichia coli

DEL VECCHIO, POMPEA GIUSEPPINA GRAZIA;BARONE, GUIDO;ROSSI, MOSE';
2006

Abstract

The thermal stability of the Aes acetyl esterase from Escherichia coli has been investigated by means of differential scanning calorimetry and circular dichroism measurements. The calorimetric curves show a denaturation temperature of 68 °C for Aes and 61 °C for the single point mutant V20D-Aes. The same values are obtained from CD denaturation curves of the two proteins recorded in both the far-UV and near-UV regions. Even if the denaturation process is irreversible and characterized by a single calorimetric peak and a single inflection point in both far- and near-UV CD curves, the overall data indicate that the process is more complex than a two-state transition. This is in line with the presence of two structural domains in the 3D model of Aes, according to homology modelling. A comparison of the thermal stability of Aes with those of the homologous thermophilic EST2 and hyperthermophilic AFEST suggests that the optimization of charge–charge interactions should not be so effective in the case of the mesophilic enzyme.
2006
Temperature-induced denaturation of Aes acetyl-esterase from Escherichia coli / DEL VECCHIO, POMPEA GIUSEPPINA GRAZIA; G., Graziano; Barone, Guido; L., Mandrich; Rossi, Mose'; G., Manco. - In: THERMOCHIMICA ACTA. - ISSN 0040-6031. - STAMPA. - 441:2(2006), pp. 144-149. [10.1016/j.tca.2005.12.006]
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11588/307102
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 5
  • ???jsp.display-item.citation.isi??? 5
social impact