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A sensitive microplate assay for lipase activity measurement using olive oil emulsion substrate: modification of the copper soap colorimetric method

2016Journal article Restricted access
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dc.contributor.authorMustafa, Ahmad
dc.contributor.authorKarmali, Amin
dc.contributor.authorAbdelmoez, Wael
dc.date.accessioned2016-12-28T14:19:06Z
dc.date.available2016-12-28T14:19:06Z
dc.date.issued2016
dc.description.abstractThe present work involves a sensitive high-throughput microtiter plate based colorimetric assay for estimating lipase activity using cupric acetate pyridine reagent (CAPR). In the first approach, three factors two levels factorial design methodology was used to evaluate the interactive effect of different parameters on the sensitivity of the assay method. The optimization study revealed that the optimum CAPR concentration was 7.5% w/v, the optimum solvent was heptane and the optimum CAPR pH was 6. In the second approach, the optimized colorimetric microplate assay was used to measure lipase activity based on enzymatic hydrolysis of olive oil emulsion substrate at 37°C and 150 rpm. The emulsion substrates were formulated by using olive oil, triton X-100 (10% v/v in pH 8) and sodium phosphate buffer of pH 8 in ratio of 1:1:1 in the case of Candida sp. lipase. While in the case of immobilized lipozyme RMIM, The emulsion substrates were formulated by using olive oil, triton X-100 (1% v/v in pH 8) and sodium phosphate buffer of pH 8 in ratio of 2:1:1. Absorbance was measured at 655 nm. The stability of this assay (in terms of colored heptane phase absorbance readings) retained more than 92.5% after 24 h at 4°C compared to the absorbance readings measured at zero time. In comparison with other lipase assay methods, beside the developed sensitivity, the reproducibility and the lower limit of detection (LOD) of the proposed method, it permits analyzing of 96 samples at one time in a 96-well microplate. Furthermore, it consumes small quantities of chemicals and unit operations.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMUSTAFA, Ahmad; KARMALI, Amin; ABDELMOEZ, Wael - A sensitive microplate assay for lípase activity measurement using olive oil emulsion substrate: modification of the cooper soap colorimetric method. Journal of Oleo Science. ISSN 1345-8957. Vol. 9, N.º 65 (2016), pp.775-784pt_PT
dc.identifier.doi10.5650/jos.ess16066pt_PT
dc.identifier.issn1345-8957
dc.identifier.urihttp://hdl.handle.net/10400.21/6680
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherJapan Oil Chemists Societypt_PT
dc.relation.publisherversionhttps://www.jstage.jst.go.jp/article/jos/65/9/65_ess16066/_pdfpt_PT
dc.subject96-well microplatept_PT
dc.subjectcolorimetric assaypt_PT
dc.subjectolive oilpt_PT
dc.subjectoleic acidpt_PT
dc.subjectCandida sp lipasept_PT
dc.subjectLipozyme RMIMpt_PT
dc.titleA sensitive microplate assay for lipase activity measurement using olive oil emulsion substrate: modification of the copper soap colorimetric methodpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage784pt_PT
oaire.citation.issue9pt_PT
oaire.citation.startPage775pt_PT
oaire.citation.titleJournal of Oleo Sciencept_PT
oaire.citation.volume65pt_PT
person.familyNameKarmali
person.familyNameAbdelmoez
person.givenNameAmin
person.givenNameWael
person.identifier.orcid0000-0003-0419-401X
person.identifier.orcid0000-0002-5370-7646
person.identifier.ridE-4787-2011
person.identifier.scopus-author-id6603778216
rcaap.rightsrestrictedAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication23975b4b-3234-4139-9675-89ea6f9a2332
relation.isAuthorOfPublication857e4d18-b0e5-45d6-8e10-e30adf61e3d3
relation.isAuthorOfPublication.latestForDiscovery23975b4b-3234-4139-9675-89ea6f9a2332

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