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An up-scalable and cost-effective methodology for isolating a polypeptide matrix metalloproteinase-9 inhibitor from Lupinus albus seeds

dc.contributor.authorMota, Joana
dc.contributor.authorFigueira, Maria E.
dc.contributor.authorFerreira, Ricardo Boavida
dc.contributor.authorLima, Ana
dc.date.accessioned2021-09-08T13:20:27Z
dc.date.available2021-09-08T13:20:27Z
dc.date.issued2021
dc.description.abstractOne of the most challenging problems with food-borne bioactive compounds is that there are commonly no cost-effective, generally recognized as safe (GRAS) methods for obtaining gram quantities of their purified forms. Here we aimed at developing a method to isolate deflamin, an oligomeric protein from lupin seeds with anti-inflammatory and anticancer activity through matrix metalloprotease (MMP)-9 inhibition. Our goal was to develop a GRAS method that could be easily up-scalable whilst maintaining deflamin’s activity. A sequential precipitation methodology was developed, using an aqueous extraction, followed by heat denaturation, acid precipitation and solubilization in ethanol. A final precipitation with 90% ethanol yielded a purified protein which was sequenced through mass spectrometry and tested for its MMP inhibitory activity using the Dyequenched (DQ) gelatin assay and the standard wound healing assay in HT29 cells. The developed method yielded a purified oligomer, which represented 0.1% (w/w) of total dry seed weight and was positively confirmed to be deflamin. It further showed to effectively reduce MMP-9 gelatinolytic activity as well as colon cancer cell migration, hence corroborating the effectiveness of our method. Overall, this is the first reported method for isolating an MMP-9 inhibitor from legume seeds, which is up-scalable to an industrial level, in a cost-effective mannerpt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMota, J.; Figueira, M.E.; Ferreira, R.B.; Lima, A. An Up-Scalable and Cost-Effective Methodology for Isolating a Polypeptide Matrix Metalloproteinase-9 Inhibitor from Lupinus albus Seeds. Foods 2021, 10, 1663pt_PT
dc.identifier.doihttps://doi.org/10.3390/ foods10071663pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.5/21765
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherMDPIpt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectMMP-9pt_PT
dc.subjectHT29pt_PT
dc.subjectGRAS-safept_PT
dc.subjectdeflaminpt_PT
dc.subjectwhite lupinpt_PT
dc.subjectnutraceuticalpt_PT
dc.titleAn up-scalable and cost-effective methodology for isolating a polypeptide matrix metalloproteinase-9 inhibitor from Lupinus albus seedspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/149830/PT
oaire.citation.titleFoodspt_PT
oaire.fundingStream3599-PPCDT
person.familyNameFerreira
person.givenNameRicardo Boavida
person.identifier86030
person.identifier.ciencia-id371D-28A0-4027
person.identifier.orcid0000-0002-5027-7564
person.identifier.scopus-author-id8285312700
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication0f32c437-6c87-47d7-998b-c97c4695806a
relation.isAuthorOfPublication.latestForDiscovery0f32c437-6c87-47d7-998b-c97c4695806a
relation.isProjectOfPublication2bea8ac8-8f47-4d13-b521-8ef0934abcec
relation.isProjectOfPublication.latestForDiscovery2bea8ac8-8f47-4d13-b521-8ef0934abcec

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