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Extraction of hemoglobin with calixarenes and biocatalysis in organic media of the complex with pseudoactivity of peroxidase

2010-01-02Journal article Open access
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dc.contributor.authorSemedo, Madga Sofia Cardoso Nobre
dc.contributor.authorKarmali, Amin
dc.contributor.authorBarata, Patrícia
dc.contributor.authorPrata, José Virgílio
dc.date.accessioned2012-03-23T16:15:36Z
dc.date.available2012-03-23T16:15:36Z
dc.date.issued2010-01-02
dc.description.abstractThe present work involves the use of p-tert-butylcalix[4,6,8]arene carboxylic acid derivatives ((t)Butyl[4,6,8]CH2COOH) for selective extraction of hemoglobin. All three calixarenes extracted hemoglobin into the organic phase, exhibiting extraction parameters higher than 0.90. Evaluation of the solvent accessible positively charged amino acid side chains of hemoglobin (PDB entry 1XZ2) revealed that there are 8 arginine, 44 lysine and 30 histidine residues on the protein surface which may be involved in the interactions with the calixarene molecules. The hemoglobin-(t)Butyl[6]CH2COOH complex had pseudoperoxidase activity which catalysed the oxidation of syringaldazine in the presence of hydrogen peroxide in organic medium containing chloroform. The effect of pH, protein and substrate concentrations on biocatalysis was investigated using the hemoglobin-(t)Butyl[6]CH2COOH complex. This complex exhibited the highest specific activity of 9.92 x 10(-2) U mg protein(-1) at an initial pH of 7.5 in organic medium. Apparent kinetic parameters (V'(max), K'(m), k'(cat) and k'(cat)/K'(m)) for the pseudoperoxidase activity were determined in organic media for different pH values from a Michaelis-Menten plot. Furthermore, the stability of the protein-calixarene complex was investigated for different initial pH values and half-life (t(1/2)) values were obtained in the range of 1.96 and 2.64 days. Hemoglobin-calixarene complex present in organic medium was recovered in fresh aqueous solutions at alkaline pH, with a recovery of pseudoperoxidase activity of over 100%. These results strongly suggest that the use of calixarene derivatives is an alternative technique for protein extraction and solubilisation in organic media for biocatalysis.por
dc.identifier.citationSemedo M C, Karmali A, Barata P D, Prata J V. Extraction of hemoglobin with calixarenes and biocatalysis in organic media of the complex with pseudoactivity of peroxidase.Journal of molecular Catalysis B-. 2010; 62 (1): 97-104.por
dc.identifier.issn1381-1177
dc.identifier.urihttp://hdl.handle.net/10400.21/1361
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherElsevier Sciencepor
dc.relation.ispartofseries1
dc.subjectCalixarenespor
dc.subjectLiquid-liquid protein extractionpor
dc.subjectHuman hemoglobinpor
dc.subjectBiocatalysis in organic mediapor
dc.subjectPseudoperoxidase activitypor
dc.subjectLiquid-extractionpor
dc.subjectRay crystal-structurespor
dc.subjectCytochrome-Cpor
dc.subjectProteinpor
dc.subjectSolventspor
dc.subjectImmobilizationpor
dc.subjectChitosanpor
dc.subjectEnzymespor
dc.subjectFormspor
dc.titleExtraction of hemoglobin with calixarenes and biocatalysis in organic media of the complex with pseudoactivity of peroxidasepor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.conferencePlaceNetherlandspor
oaire.citation.endPage104por
oaire.citation.issue62por
oaire.citation.startPage97por
oaire.citation.titleJournal of molecular Catalysis B-por
person.familyNameSemedo
person.familyNameKarmali
person.familyNameBarata
person.familyNamePrata
person.givenNameMagda
person.givenNameAmin
person.givenNamePatrícia
person.givenNameJosé Virgílio
person.identifier1022083
person.identifier1090399
person.identifier191765
person.identifier.ciencia-id9C11-BD02-C77F
person.identifier.ciencia-idC019-9F30-0F99
person.identifier.ciencia-idC11A-9130-8981
person.identifier.orcid0000-0001-6330-2899
person.identifier.orcid0000-0003-0419-401X
person.identifier.orcid0000-0001-8274-633X
person.identifier.orcid0000-0002-9068-2847
person.identifier.ridE-4787-2011
person.identifier.ridB-7214-2008
person.identifier.scopus-author-id57000381900
person.identifier.scopus-author-id6603778216
person.identifier.scopus-author-id8968446000
person.identifier.scopus-author-id9276778400
rcaap.rightsrestrictedAccesspor
rcaap.typearticlepor
relation.isAuthorOfPublication73e398bc-a0e8-420d-81eb-34159398838d
relation.isAuthorOfPublication23975b4b-3234-4139-9675-89ea6f9a2332
relation.isAuthorOfPublication4ab8a01f-c11d-4b3b-b0cf-b21e5b634170
relation.isAuthorOfPublication15e229f3-a619-4931-87eb-799091dc1601
relation.isAuthorOfPublication.latestForDiscovery73e398bc-a0e8-420d-81eb-34159398838d

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