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Shear-stress-mediated refolding of proteins from aggregates and inclusion bodies.

ABSTRACT: Recombinant protein overexpression of large proteins in bacteria often results in insoluble and misfolded proteins directed to inclusion bodies. We report the application of shear stress in micrometer-wide, thin fluid films to refold boiled hen egg white lysozyme, recombinant hen egg white lysozyme, and recombinant caveolin-1. Furthermore, the approach allowed refolding of a much larger protein, cAMP-dependent protein kinase A (PKA). The reported methods require only minutes, which is more than 100 times faster than conventional overnight dialysis. This rapid refolding technique could significantly shorten times, lower costs, and reduce waste streams associated with protein expression for a wide range of industrial and research applications.

SUBMITTER: Yuan TZ

PROVIDER: S-EPMC4388321 | biostudies-literature | 2015 Feb

REPOSITORIES: biostudies-literature

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Shear-stress-mediated refolding of proteins from aggregates and inclusion bodies.

Yuan Tom Z TZ Ormonde Callum F G CF Kudlacek Stephan T ST Kunche Sameeran S Smith Joshua N JN Brown William A WA Pugliese Kaitlin M KM Olsen Tivoli J TJ Iftikhar Mariam M Raston Colin L CL Weiss Gregory A GA

Chembiochem : a European journal of chemical biology 20150123 3

Recombinant protein overexpression of large proteins in bacteria often results in insoluble and misfolded proteins directed to inclusion bodies. We report the application of shear stress in micrometer-wide, thin fluid films to refold boiled hen egg white lysozyme, recombinant hen egg white lysozyme, and recombinant caveolin-1. Furthermore, the approach allowed refolding of a much larger protein, cAMP-dependent protein kinase A (PKA). The reported methods require only minutes, which is more than ...[more]

PMID: 25620679

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