Effect of pressure on refolding of recombinant pentameric cholera toxin B
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2014
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Journal of Biotechnology
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The production of recombinant proteins is an essential tool for the expansion of modern biological
research and biotechnology. The expression of heterologous proteins in Escherichia coli often results
in an incomplete folding process that leads to the accumulation of inclusion bodies (IB), aggregates
that hold a certain degree of native-like secondary structure. High hydrostatic pressure (HHP) impairs
intermolecular hydrophobic and electrostatic interactions, leading to dissociation of aggregates under
non-denaturing conditions and is therefore a useful tool to solubilize proteins for posterior refolding.
Cholera toxin (CT) is composed of a non-toxic pentamer of B subunits (CTB), a useful adjuvant in vaccines, and a toxic subunit A (CTA). We studied the process of refolding of CTB using HHP. HHP was shown
to be effective for dissociation of CTB monomers from IB. Posterior incubation at atmospheric pressure
of concentrated CTB (1 mg/ml) is necessary for the association of the monomers. Pentameric CTB was
obtained when suspensions of CTB IB were compressed at 2.4 kbar for 16 h in the presence of Tween 20
and incubated at 1 bar for 120 h. Soluble and biologically active pentameric CTB was obtained, with a
yield of 213 mg CTB/liter of culture. The experience gained in this study can be important to improve the
refolding of proteins with quaternary structure.
Como referenciar
RODRIGUES, D.; FARINHA-ARCIERI, L.E.; VENTURA, A.M.; CHURA-CHAMBI, R.M.; MALAVASI, R.V.; LEMKE, L.S.; GUIMARAES, J.S.; HO, P.L.; MORGANTI, L. Effect of pressure on refolding of recombinant pentameric cholera toxin B. Journal of Biotechnology, v. 173, p. 98-105, 2014. DOI: 10.1016/j.jbiotec.2013.12.006. Disponível em: http://repositorio.ipen.br/handle/123456789/8951. Acesso em: 12 May 2024.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.