title: Mixed ion exchange supports as useful ion exchangers for protein purification: purification of penicillin G acylase from Escherichia coli.
abstract: A support having similar amounts of carboxymethyl and amino groups has been prepared and evaluated as an ion exchanger. It has been found that this support was able to adsorb a high amount of protein from a crude extract of proteins (approximately 55%) at pH 5. Moreover, it was able to adsorb approximately 60% of the protein that did not become adsorbed on supports bearing just one kind of ionic groups. The use of divalent cations reinforced the adsorption of proteins on these supports. These results suggest that the adsorption of proteins on supports bearing almost neutral charge is not driven by the existence of opposite charges between the adsorbent and the biomacromolecule but just by the possibility of forming a high number of enzyme-support ionic bonds. This support has been used to purify the enzyme penicillin G acylase (PGA) from Escherichia coli. PGA was not significantly adsorbed at any pH value on either amino- or carboxyl-activated supports, while it can be fully adsorbed at pH 5 on this new carboxyl-amino matrix. Thus, we have been able to almost fully purify PGA from crude extracts with a very high yield by using these new supports.
authors.affiliation: Departamento de BiocatAlisis, Instituto de CatAlisis y Petroleoquimíca, Consejo Superior de Invesigaciones Cientificas, Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain.
authors.forename: Manuel
authors.initials: M
authors.lastname: Fuentes
authors.forename: Pilar
authors.initials: P
authors.lastname: Batalla
authors.forename: Valeria
authors.initials: V
authors.lastname: Grazu
authors.forename: Benevides C C
authors.initials: BC
authors.lastname: Pessela
authors.forename: Cesar
authors.initials: C
authors.lastname: Mateo
authors.forename: Tamara
authors.initials: T
authors.lastname: Montes
authors.forename: Juan A
authors.initials: JA
authors.lastname: Hermoso
authors.forename: Jose M
authors.initials: JM
authors.lastname: Guisan
authors.forename: Roberto
authors.initials: R
authors.lastname: Fernandez-Lafuente
mesh_terms: D000327:Adsorption; D029968:Escherichia coli Proteins / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007474:Ion Exchange; D010399:Penicillin Amidase / Q000302:isolation & purification*; D011506:Proteins / Q000302:isolation & purification
chemical_list: D029968:Escherichia coli Proteins; D011506:Proteins; D010399:Penicillin Amidase