Archaea

Archaea / 2005 / Article

Research Article | Open Access

Volume 1 |Article ID 543789 | https://doi.org/10.1155/2005/543789

Anna Scotto D’Abusco, Rita Casadio, Gianluca Tasco, Laura Giangiacomo, Anna Giartosio, Valentina Calamia, Stefania Di Marco, Roberta Chiaraluce, Valerio Consalvi, Roberto Scandurra, Laura Politi, "Oligomerization of Sulfolobus solfataricus signature amidase is promoted by acidic pH and high temperature", Archaea, vol. 1, Article ID 543789, 13 pages, 2005. https://doi.org/10.1155/2005/543789

Oligomerization of Sulfolobus solfataricus signature amidase is promoted by acidic pH and high temperature

Received04 Jul 2005
Accepted13 Sep 2005

Abstract

The recombinant amidase from the hyperthermophylic archaeon Sulfolobus solfataricus (SSAM) a signature amidase, was cloned, purified and characterized. The enzyme is active on a large number of aliphatic and aromatic amides over the temperature range 60–95 °C and at pH values between 4.0 and 9.5, with an optimum at pH 5.0. The recombinant enzyme is in the form of a dimer of about 110 kD that reversibly associates into an octamer in a pH-dependent reaction. The pH dependence of the state of association was studied using gel permeation chromatography, analytical ultracentrifugation and dynamic light scattering techniques.pH 7.0 all three techniques show the presence of two species, in about equal amounts, which is compatible with the existence of a dimeric and an octameric form. In decreasing pH, the dimers formed the octameric species and in increasing pH, the octameric species was converted to dimers. Above pH 8.0, only dimers were present, below pH 3.0 only octamers were present. The association of dimers into octamers decreased in non-polar solvents and increased with temperature. A mutant (Y41C) was obtained that did not show this behavior.

Copyright © 2005 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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