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Pseudomonas aeruginosa amidase: Aggregation in recombinant Escherichia coli

2011-07Journal article Open access
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dc.contributor.authorBorges, Patrícia
dc.contributor.authorPacheco, Rita
dc.contributor.authorKarmali, Amin
dc.date.accessioned2013-02-16T17:44:11Z
dc.date.available2013-02-16T17:44:11Z
dc.date.issued2011-07
dc.description.abstractThe effect of cultivation parameters such as temperature incubation, IPTG induction and ethanol shock on the production of Pseudomonasaeruginosa amidase (E.C.3.5.1.4) in a recombinant Escherichia coli strain in LB ampicillin culture medium was investigated. The highest yield of solubleamidase, relatively to other proteins, was obtained in the condition at 37 degrees C using 0.40 mM IPTG to induce growth, with ethanol. Our results demonstrate the formation of insoluble aggregates containing amidase, which was biologically active, in all tested growth conditions. Addition of ethanol at 25 degrees C in the culture medium improved amidase yield, which quantitatively aggregated in a biologically active form and exhibited in all conditions an increased specific activity relatively to the soluble form of the enzyme. Non-denaturing solubilization of the aggregated amidase was successfully achieved using L-arginine. The aggregates obtained from conditions at 37 degrees C by Furier transform infrared spectroscopy (FTIR) analysis demonstrated a lower content of intermolecular interactions, which facilitated the solubilization step applying non-denaturing conditions. The higher interactions exhibited in aggregates obtained at suboptimal conditions compromised the solubilization yield. This work provides an approach for the characterization and solubilization of novel reported biologically active aggregates of this amidase.por
dc.identifier.citationBORGES, Patrícia; PACHECO, Rita; KARMALI, Amin - Pseudomonas aeruginosa amidase: Aggregation in recombinant Escherichia coli. Biotechnology Journal. ISSN 1860-6768. Vol. 6, n.º 7 (2011) p. 888-897.por
dc.identifier.issn1860-6768
dc.identifier.urihttp://hdl.handle.net/10400.21/2231
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherWiley-Blackwellpor
dc.subjectEscherichia Colipor
dc.subjectFTIRpor
dc.subjectProtein aggregatespor
dc.subjectRecombinant amidasepor
dc.subjectSolubilizationpor
dc.subjectTransform infrared-spectroscopypor
dc.subjectBacterial inclusion-bodiespor
dc.subjectActive-site nucleophilepor
dc.subjectFT-IR spectroscopypor
dc.subjectHydromaxic acidspor
dc.subjectMonoclonal-antibodiespor
dc.subjectSecondary structurepor
dc.subjectL-Argininepor
dc.subjectProteinspor
dc.subjectSolubilizationpor
dc.titlePseudomonas aeruginosa amidase: Aggregation in recombinant Escherichia colipor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.conferencePlaceMaldenpor
oaire.citation.endPage897por
oaire.citation.issue7por
oaire.citation.startPage888por
oaire.citation.titleBiotechnology Journalpor
oaire.citation.volume6por
person.familyNamePacheco
person.familyNameKarmali
person.givenNameRita
person.givenNameAmin
person.identifier1022927
person.identifier.ciencia-id9F13-D310-B5A3
person.identifier.orcid0000-0001-5192-3006
person.identifier.orcid0000-0003-0419-401X
person.identifier.ridC-3062-2012
person.identifier.ridE-4787-2011
person.identifier.scopus-author-id8853708300
person.identifier.scopus-author-id6603778216
rcaap.rightsrestrictedAccesspor
rcaap.typearticlepor
relation.isAuthorOfPublicationd6907c0a-3f8f-431c-acf2-0b257692f0d4
relation.isAuthorOfPublication23975b4b-3234-4139-9675-89ea6f9a2332
relation.isAuthorOfPublication.latestForDiscovery23975b4b-3234-4139-9675-89ea6f9a2332

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