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Micro-analytical GO/HRP bioreactor for glucose determination and bioprocess monitoring

2005-04-15Journal article Open access
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dc.contributor.authorVojinović, V.
dc.contributor.authorCalado, Cecília
dc.contributor.authorSilva, A. I.
dc.contributor.authorMateus, M.
dc.contributor.authorCabral, J. M. S.
dc.contributor.authorFonseca, Luís P. P.
dc.date.accessioned2020-09-23T10:44:11Z
dc.date.available2020-09-23T10:44:11Z
dc.date.issued2005-04-15
dc.description.abstractA bi-enzymatic micro-analytical bioreactor integrated in a FIA system for glucose measurements is described. Its robustness and small dimensions (working volume of about 70 μl containing approximately 1.2 mg GO and 0.26 mg HRP) make it easy to operate. The column is based on immobilisation of glucose oxidase (GO) and horseradish peroxidase (HRP) on alkylamine controlled pore glass (CPG) beads. The column has excellent shelf life (no significant loss of activity after 1 year if kept at 4 °C), and a very high operational stability that was demonstrated through extensive usage for glucose determinations over 1 year period during which the column retained almost all of its activity. More importantly, this operational stability allows glucose monitoring in the culture media without a decay of signal over the experiment time and consequently no signal correction or re-calibration is needed. This high operational stability was also confirmed by continuous glucose conversion with 30% activity loss after converting quantity of glucose equivalent to 21600 FIA injections of 20 μl with 1.7 mM glucose. Such good performance is a result of an optimised immobilisation method and moreover of the implementation of in situ enzyme stabilisation strategy which consisted on promoting the instantaneous H2O2 consumption produced by the GO. This strategy has the additional advantage of allowing concomitant assay of the H2O2 based on the HAP catalysed co-oxidation of phenol-4-sulphonic acid (PSA) in the presence of 4-aminoantipyrine (4-AAP). The glucose measurements are reproducible with high precision against the standard HPLC method. Linear range and sensitivity depend on sample injection volume; the upper limit is about 1.1 g/l. Lower detection limit is 10 mg/l. The column performance has been validated for E. coli and S. cerevisiae fermentation monitoring, and glucose measurements in an animal cell culture (rat Langerhans islets).pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationVOJINOVIĆ, V.; [et al] – Micro-analytical GO/HRP bioreactor for glucose determination and bioprocess monitoring. Biosensors and Bioelectronics. ISSN 0956-5663. Vol. 20, N.º 10 (2005), pp. 1955-1961pt_PT
dc.identifier.doi10.1016/j.bios.2004.08.015pt_PT
dc.identifier.issn0956-5663
dc.identifier.urihttp://hdl.handle.net/10400.21/12252
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relation.publisherversionhttps://reader.elsevier.com/reader/sd/pii/S0956566304003811?token=C86123A63DEFBCA31AAFD6CB31852775CD6210C1C72569CDF432F2DFC5E2D19FD353A23158FB0D557D1B4C8A662B93E9pt_PT
dc.subjectBiosensorpt_PT
dc.subjectGlucosept_PT
dc.subjectMonitoringpt_PT
dc.subjectBi-enzymatic columnpt_PT
dc.subjectHorseradish peroxidasept_PT
dc.subjectGlucose oxidasept_PT
dc.titleMicro-analytical GO/HRP bioreactor for glucose determination and bioprocess monitoringpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBD%2F5495%2F2001/PT
oaire.citation.endPage1961pt_PT
oaire.citation.issue10pt_PT
oaire.citation.startPage1955pt_PT
oaire.citation.titleBiosensors and Bioelectronicspt_PT
oaire.citation.volume20pt_PT
oaire.fundingStreamSFRH
person.familyNameCalado
person.familyNameP. Fonseca
person.givenNameCecília
person.givenNameLuis
person.identifier130332
person.identifier.ciencia-id9418-E320-3177
person.identifier.ciencia-id951D-DF38-3397
person.identifier.orcid0000-0002-5264-9755
person.identifier.orcid0000-0001-8429-6977
person.identifier.ridE-2102-2014
person.identifier.ridA-4228-2013
person.identifier.scopus-author-id6603163260
person.identifier.scopus-author-id55916288400
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublicatione8577257-c64c-4481-9b2b-940fedb360cc
relation.isAuthorOfPublication5c533551-5113-4c0a-93f1-9c50cbd796bc
relation.isAuthorOfPublication.latestForDiscoverye8577257-c64c-4481-9b2b-940fedb360cc
relation.isProjectOfPublicationcd5f22d0-bebd-4289-9831-18e599556e32
relation.isProjectOfPublication.latestForDiscoverycd5f22d0-bebd-4289-9831-18e599556e32

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