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dc.contributor.authorHassan, Rabeay Y A
dc.contributor.authorBilitewski, Ursula
dc.date.accessioned2012-03-22T14:02:04Z
dc.date.available2012-03-22T14:02:04Z
dc.date.issued2011-12-01
dc.identifier.citationA viability assay for Candida albicans based on the electron transfer mediator 2,6-dichlorophenolindophenol. 2011, 419 (1):26-32 Anal. Biochem.en_GB
dc.identifier.issn1096-0309
dc.identifier.pmid21864496
dc.identifier.doi10.1016/j.ab.2011.07.025
dc.identifier.urihttp://hdl.handle.net/10033/216297
dc.description.abstractCandida albicans is an opportunistic fungal pathogen with comparably high respiratory activity. Thus, we established a viability test based on 2,6-dichlorophenolindophenol (DCIP), a membrane-permeable electron transfer agent. NADH dehydrogenases catalyze the reduction of DCIP by NADH, and the enzymatic activity can be determined either electrochemically via oxidation reactions of DCIP or photometrically. Among the specific respiratory chain inhibitors, only the complex I inhibitor rotenone decreased the DCIP signal from C. albicans, leaving residual activity of approximately 30%. Thus, the DCIP-reducing activity of C. albicans was largely dependent on complex I activity. C. albicans is closely related to the complex I-negative yeast Saccharomyces cerevisiae, which had previously been used in DCIP viability assays. Via comparative studies, in which we included the pathogenic complex I-negative yeast Candida glabrata, we could define assay conditions that allow a distinction of complex I-negative and -positive organisms. Basal levels of DCIP turnover by S.cerevisiae and C. glabrata were only 30% of those obtained from C. albicans but could be increased to the C. albicans level by adding glucose. No significant increases were observed with galactose. DCIP reduction rates from C. albicans were not further increased by any carbon source.
dc.language.isoenen
dc.rightsArchived with thanks to Analytical biochemistryen_GB
dc.subject.mesh2,6-Dichloroindophenolen_GB
dc.subject.meshCandida albicansen_GB
dc.subject.meshCandida glabrataen_GB
dc.subject.meshCatalysisen_GB
dc.subject.meshColoren_GB
dc.subject.meshElectrochemistryen_GB
dc.subject.meshElectron Transporten_GB
dc.subject.meshEnzyme Activationen_GB
dc.subject.meshFungal Proteinsen_GB
dc.subject.meshGalactoseen_GB
dc.subject.meshGlucoseen_GB
dc.subject.meshMicrobial Viabilityen_GB
dc.subject.meshMicrobiological Techniquesen_GB
dc.subject.meshNADen_GB
dc.subject.meshNADH Dehydrogenaseen_GB
dc.subject.meshOxidation-Reductionen_GB
dc.subject.meshOxygenen_GB
dc.subject.meshRotenoneen_GB
dc.subject.meshSaccharomyces cerevisiaeen_GB
dc.subject.meshSpectrophotometryen_GB
dc.titleA viability assay for Candida albicans based on the electron transfer mediator 2,6-dichlorophenolindophenol.en
dc.typeArticleen
dc.contributor.departmentBiological Systems Analysis Group, Helmholtz Centre for Infection Research, 38124 Braunschweig, Germany.en_GB
dc.identifier.journalAnalytical biochemistryen_GB
refterms.dateFOA2018-06-13T01:19:55Z
html.description.abstractCandida albicans is an opportunistic fungal pathogen with comparably high respiratory activity. Thus, we established a viability test based on 2,6-dichlorophenolindophenol (DCIP), a membrane-permeable electron transfer agent. NADH dehydrogenases catalyze the reduction of DCIP by NADH, and the enzymatic activity can be determined either electrochemically via oxidation reactions of DCIP or photometrically. Among the specific respiratory chain inhibitors, only the complex I inhibitor rotenone decreased the DCIP signal from C. albicans, leaving residual activity of approximately 30%. Thus, the DCIP-reducing activity of C. albicans was largely dependent on complex I activity. C. albicans is closely related to the complex I-negative yeast Saccharomyces cerevisiae, which had previously been used in DCIP viability assays. Via comparative studies, in which we included the pathogenic complex I-negative yeast Candida glabrata, we could define assay conditions that allow a distinction of complex I-negative and -positive organisms. Basal levels of DCIP turnover by S.cerevisiae and C. glabrata were only 30% of those obtained from C. albicans but could be increased to the C. albicans level by adding glucose. No significant increases were observed with galactose. DCIP reduction rates from C. albicans were not further increased by any carbon source.


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