Please use this identifier to cite or link to this item: http://hdl.handle.net/1822/22630

TitleProtein disulphide isomerase-induced refolding of sonochemically prepared Ribonuclease A microspheres
Author(s)Fernandes, Margarida M.
Silva, Raquel
Ferreira, Helena
Donelli, Ilaria
Freddi, G.
Paulo, Artur Cavaco
KeywordsProtein disulphide isomerase
Ribonuclease A
Ultrasound
Microspheres
Activity recovery
Protein release
Protein folding
Issue date2012
PublisherElsevier
JournalJournal of Biotechnology
Abstract(s)The present communication describes for the first time the development of Ribonuclease A (RNase A) microspheres using the sonochemical method followed by an enzymatic treatment with protein disulphide isomerase (PDI). Ultrasound application induced changes on the protein physicochemical and biological properties: the enzymatic activity of RNase A was decreased in 35% and the free thiol groups content was significantly increased, probably due to the breakage of protein disulphide bonds and assembly of RNase A monomers. The deconvolution of amide I band, from Fourier Transform Infrared Spectroscopy, showed that the secondary structure of RNase A was slightly changed after microspherization. The PDI application on microspheres promoted the recovery of RNase A biological activity and induced the release of active protein into solution in its native state. These results were promoted by different states of PDI active site: oxidized and reduced, respectively. The PDI aptitude to catalyze the refolding of a protein substrate in the form of spheres is here reported.
TypeArticle
URIhttp://hdl.handle.net/1822/22630
DOI10.1016/j.jbiotec.2012.02.016
ISSN0168-1656
Publisher version10.1016/j.jbiotec.2012.02.016
Peer-Reviewedyes
AccessOpen access
Appears in Collections:DET/2C2T - Artigos em revistas internacionais com arbitragem científica

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