Please use this identifier to cite or link to this item: http://hdl.handle.net/123456789/234
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dc.contributor.authorGarg, Lalit C-
dc.date.accessioned2014-11-26T09:19:44Z-
dc.date.available2014-11-26T09:19:44Z-
dc.date.issued2012-01-
dc.identifier.urihttp://hdl.handle.net/123456789/234-
dc.description.abstractInclusion bodies of recombinant human growth hormone (r-hGH) were isolated from Escherichia coli, enriched and solubilized in 100mM Tris buffer containing 6M n-propanol and 2M urea. Around 4 mg/ml of r-hGH from inclusion bodies were solubilized in 6M n-propanol-based buffer containing 2M urea. Existence of native-like secondary structure of r-hGH in 6M n-propanol solution was confirmed by CD and fluorescence spectra. Solubilized r-hGH was subsequently refolded by pulsatile dilution, purified to homogeneity and found to be functionally active. Tris buffer containing 6M n-propanol and 2M urea also effectively solubilized a number of proteins expressed as inclusion bodies in E. coli. Mild solubilization of inclusion body proteins, chaotropic effect of n-propanol at high concentration and kosmotropic effect at lower concentration helped in improved refolding of the solubilized protein. Around 40% of the r-hGH in the form of inclusion body aggregates was refolded into bioactive form while using n-propanol as solubilization agent. Solubilization with 6M n-propanol solution thus can be a viable alternative for achieving high throughput recovery of bioactive protein from inclusion bodies of E. coli.en_US
dc.publisherElsevier Incen_US
dc.subjectProtein Expressionen_US
dc.subjectGene regulationen_US
dc.titleSolubilization of inclusion body proteins using n-propanol and its refolding into bioactive formen_US
dc.contributor.coauthorSingh, Surinder M-
dc.contributor.coauthorSharma, Aparna-
dc.contributor.coauthorUpadhyay, Arun K-
dc.contributor.coauthorSingh, Anupam-
dc.contributor.coauthorPanda, Amulya K-
dc.keywordRecombinant human growth hormone, Escherichia coli, Inclusion bodies, n-Propanol, Solubilization, Refolding and purificationen_US
dc.journalProtein Expression and Purificationen_US
dc.volumeno81en_US
dc.issueno1en_US
dc.pages75-82en_US
Appears in Collections:Gene Regulation, Publications

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