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A novel prokaryotic expression system for biosynthesis of recombinant human membrane-bound catechol-O-methyltransferase

2011-11-10Journal article Restricted access
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dc.contributor.authorPedro, Augusto
dc.contributor.authorBonifácio, Maria João
dc.contributor.authorQueiroz, J. A.
dc.contributor.authorMaia, C J
dc.contributor.authorPassarinha, L A
dc.date.accessioned2019-11-27T09:34:51Z
dc.date.available2019-11-27T09:34:51Z
dc.date.issued2011-11-10
dc.description.abstractMembrane proteins constitute 20-30% of all proteins encoded by the genome of various organisms. While large amounts of purified proteins are required for pharmaceutical and crystallization attempts, there is an unmet need for the development of novel heterologous membrane protein overexpression systems. Specifically, we tested the application of Brevibacillus choshinensis cells for the biosynthesis of human membrane bound catechol-O-methyltransferase (hMBCOMT). In terms of the upstream stage moderate to high expression was obtained for complex media formulation with a value near 45 nmol/h/mg for hMBCOMT specific activity achieved at 20 h culture with 37°C and 250 rpm. Subsequently, the efficiency for reconstitution of hMBCOMT is markedly null in the presence of ionic detergents, such as sodium dodecyl sulphate (SDS). In general, for non-ionic and zwiterionic detergents, until a detergent critic micellar concentration (CMC) of 1.0 mM, hMBCOMT shows more biological activity at lower detergent concentrations while for detergent CMC higher than 1 mM, higher detergent concentrations seem to be ideal for hMBCOMT solubilization. Indeed, from the detergents tested, the non-ionic digitonin at 0.5% (w/v) appears to be the most suitable for hMBCOMT solubilization.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1016/j.jbiotec.2011.08.022pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.6/7623
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.subjectMembrane Proteinspt_PT
dc.subjectRecombinant Proteinspt_PT
dc.subjectHuman MBCOMTpt_PT
dc.subjectHeterologous protein productionpt_PT
dc.subjectDetergentspt_PT
dc.subjectParkinson diseasept_PT
dc.titleA novel prokaryotic expression system for biosynthesis of recombinant human membrane-bound catechol-O-methyltransferasept_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage146pt_PT
oaire.citation.issue2pt_PT
oaire.citation.startPage141-6pt_PT
oaire.citation.titleJournal of Biotechnologypt_PT
oaire.citation.volume156pt_PT
person.familyNameQuaresma Henriques Pedro
person.familyNameQueiroz
person.familyNameMaia Baptista
person.familyNamePaulino Passarinha
person.givenNameAugusto
person.givenNameJoão
person.givenNameCláudio Jorge
person.givenNameLuís António
person.identifierhttps://scholar.google.com/citations?user=oer44UoAAAAJ&hl=pt-PT&oi=ao
person.identifierhttps://scholar.google.pt/citations?user=hFKOd_QAAAAJ&hl=pt-PT&oi=ao
person.identifierhttps://www.researchgate.net/profile/Luis_Passarinha
person.identifier.ciencia-id7111-B2BE-336D
person.identifier.ciencia-id931E-B66D-E341
person.identifier.ciencia-id5115-FCBD-A31F
person.identifier.ciencia-id621B-74BF-F9A7
person.identifier.orcid0000-0001-8194-6022
person.identifier.orcid0000-0002-3096-8325
person.identifier.orcid0000-0002-5658-5445
person.identifier.orcid0000-0001-6910-7576
person.identifier.ridL-3104-2014
person.identifier.scopus-author-id54080426100
person.identifier.scopus-author-id7003705645
person.identifier.scopus-author-id23097426600
person.identifier.scopus-author-id12445439600
rcaap.embargofctCopyright cedido à editora no momento da publicação.pt_PT
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
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relation.isAuthorOfPublicationc798c033-c160-4f99-a8ad-04416923943f
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relation.isAuthorOfPublication.latestForDiscoverya431a31c-1ae0-43e9-8d84-4e9caca2ad92

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