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dc.creatorMacinkovic, Igor S.
dc.creatorAbughren, Mohamed
dc.creatorMrkić, Ivan
dc.creatorGrozdanovic, Milica M.
dc.creatorProdanović, Radivoje
dc.creatorGavrović-Jankulović, Marija
dc.date.accessioned2018-11-22T00:24:43Z
dc.date.available2018-11-22T00:24:43Z
dc.date.issued2013
dc.identifier.issn0168-1656
dc.identifier.urihttp://cherry.chem.bg.ac.rs/handle/123456789/1449
dc.description.abstractHigh levels of recombinant protein expression can lead to the formation of insoluble inclusion bodies. These complex aggregates are commonly solubilized in strong denaturants, such as 6-8 M urea, although, if possible, solubilization under milder conditions could facilitate subsequent refolding and purification of bioactive proteins. Commercially available GST-tag assays are designed for quantitative measurement of GST activity under native conditions. GST fusion proteins accumulated in inclusion bodies are considered to be undetectable by such assays. In this work, solubilization of recombinantly produced proteins was performed in 4 M urea. The activity of rGST was assayed in 2 M urea and it was shown that rGST preserves 85% of its activity under such denaturing conditions. A colorimetric GST activity assay with 1-chloro-2, 4-dinitrobenzene (CDNB) was examined for use in rapid detection of expression targeted to inclusion bodies and for the identification of inclusion body proteins which can be solubilized in low concentrations of chaotropic agents. Applicability of the assay was evaluated by tracking protein expression of two GST-fused allergens of biopharmaceutical value in E. coli, GST-Der p 2 and GST-Mus a 5, both targeted to inclusion bodies. (C) 2013 Elsevier B.V. All rights reserved.en
dc.publisherElsevier Science Bv, Amsterdam
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/172049/RS//
dc.rightsrestrictedAccess
dc.sourceJournal of Biotechnology
dc.subjectGSTen
dc.subjectGST-tagged proteinsen
dc.subjectInclusion bodiesen
dc.subjectUreaen
dc.titleEmployment of colorimetric enzyme assay for monitoring expression and solubility of GST fusion proteins targeted to inclusion bodiesen
dc.typearticle
dc.rights.licenseARR
dcterms.abstractМркић, Иван; Aбугхрен, Мохамед; Гавровић-Јанкуловић, Марија; Гроздановиц, Милица М.; Продановић, Радивоје; Мацинковиц, Игор С.;
dc.citation.volume168
dc.citation.issue4
dc.citation.spage506
dc.citation.epage510
dc.identifier.wos000327843200030
dc.identifier.doi10.1016/j.jbiotec.2013.09.019
dc.citation.other168(4): 506-510
dc.citation.rankM21
dc.identifier.pmid24100211
dc.type.versionpublishedVersionen
dc.identifier.scopus2-s2.0-84888826710
dc.identifier.rcubKon_2569


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