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dc.contributor.authorUsa Boonyuenen_US
dc.contributor.authorKamoltip Promnaresen_US
dc.contributor.authorSuwapat Junkreeen_US
dc.contributor.authorNichloas P.J. Dayen_US
dc.contributor.authorMallika Imwongen_US
dc.contributor.otherMahidol Universityen_US
dc.contributor.otherPrince of Songkla Universityen_US
dc.contributor.otherChurchill Hospitalen_US
dc.date.accessioned2018-11-23T09:49:11Z-
dc.date.available2018-11-23T09:49:11Z-
dc.date.issued2015-01-01en_US
dc.identifier.citationProtein Expression and Purification. Vol.107, (2015), 68-75en_US
dc.identifier.issn10465928en_US
dc.identifier.other2-s2.0-84918521617en_US
dc.identifier.urihttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84918521617&origin=inwarden_US
dc.identifier.urihttp://repository.li.mahidol.ac.th/dspace/handle/123456789/35584-
dc.description.abstract© 2014 The Authors. Published by Elsevier Inc. Human liver carboxylesterase 1 (CES1) plays a critical role in the hydrolysis of various ester- and amide-containing molecules, including active metabolites, drugs and prodrugs. However, it has been problematic to express recombinant CES1 in bacterial expression systems due to low solubility, with the CES1 protein being mainly expressed in inclusion bodies, accompanied by insufficient purity issues. In this study, we report an efficient in vitro method for refolding recombinant CES1 from inclusion bodies. A one-step purification with an immobilized-metal affinity column was utilized to purify His-tagged recombinant CES1. Conveniently, both denaturant and imidazole can be removed while the enzyme is refolded via buffer exchange, a dilution method. We show that the refolding of recombinant CES1 was successful in Tris-HCl at pH 7.5 containing a combination of 1% glycerol and 2 mM β-mercaptoethanol, whereas a mixture of other additives (trehalose, sorbitol and sucrose) and β-mercaptoethanol failed to recover a functional protein. His-tagged recombinant CES1 retains its biological activity after refolding and can be used directly without removing the fusion tag. Altogether, our results provide an alternative method for obtaining a substantial amount of functionally active protein, which is advantageous for further investigations such as structural and functional studies.en_US
dc.rightsMahidol Universityen_US
dc.source.urihttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84918521617&origin=inwarden_US
dc.subjectBiochemistry, Genetics and Molecular Biologyen_US
dc.titleEfficient in vitro refolding and functional characterization of recombinant human liver carboxylesterase (CES1) expressed in E. colien_US
dc.typeArticleen_US
dc.rights.holderSCOPUSen_US
dc.identifier.doi10.1016/j.pep.2014.11.006en_US
Appears in Collections:Scopus 2011-2015

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