ROSA MARIA CHURA CHAMBI

ROSA MARIA CHURA CHAMBI

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Investigation on solubilization protocols in the refolding of the thiredoxin TsnC from Xylella fastidiosa by high hydrostatic pressure approach
2015 - LEMKE, LAURA S.; CHURA-CHAMBI, ROSA M.; RODRIGUES, DANIELLA; CUSSIOL, JOSE R.R.; MALAVASI, NATALIA V.; ALEGRIA, THIAGO G.P.; SOARES NETTO, LUIS E.; MORGANTI, LIGIA
Effect of pressure on refolding of recombinant pentameric cholera toxin B
2014 - 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.
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.
Application of high hydrostatic pressure for recovery of biologically active cruzain from recombinant inclusion bodies
2011 - CHAMBI, ROSA M.C.; LEMKE, LAURA S.; VALLEJO, NATALIA M.; JULIANO NETO, LUIZ; MORGANTI, LIGIA
The effect of temperature on protein refolding at high pressure enhanced green fluorescent protein as a model
2014 - MALAVASI, N.V.; CORDEIRO, Y.; RODRIGUES, D.; CHURA-CHAMBI, R.M.; LEMKE, L.S.; MORGANTI, L.
An analysis of the factors that affect the dissociation of inclusion bodies and the refolding of endostatin under high pressure
2013 - CHURA-CHAMBI, R.M.; CORDEIRO, Y.; MALAVASI, N.V.; LEMKE, L.S.; RODRIGUES, D.; MORGANTI, L.
Refolding of endostatin from inclusion bodies using high hydrostatic pressure
2008 - CHURA-CHAMBI, ROSA M.; GENOVA, LUIS A.; AFFONSO, REGINA; MORGANTI, LIGIA
High hydrostatic pressure was used for concomitant solubilization and refolding of insoluble endostatin (ES) aggregated as inclusion bodies (IBs). High hydrostatic pressure (200 MPa or 2 kbar) was applied in combination with nondenaturing concentrations of guanidine hydrochloride. High levels of correctly folded ES (90 mg/L culture) were obtained after optimization/standardization of the procedure by applying pressures of 200 MPa for 16 h in 1.5 M guanidine hydrochloride/0.5 mM oxidized glutathione and reduced glutathione. Refolded ES was purified by affinity chromatography on a heparin column and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, size exclusion HPLC, circular dichroism, and intrinsic fluorescence. We demonstrated that high pressure can successfully convert insoluble IBs of ES expressed in Escherichia coli into an ES preparation with native tertiary structure and full biological activity.
Refolding on the recombinant protein OmpA70 from Leptospira interrogans from inclusion bodies using high hydrostatic pressure and partial characterization of its immunological properties
2010 - FRAGA, TATIANA R.; CHURA-CHAMBI, ROSA M.; GONCALEZ, AMANE P.; MORAIS, ZENAIDE M.; VASCONCELLOS, SILVIO A.; MORGANTI, LIGIA; MARTINS, ELIZABETH A.L.
Refolding by high pressure of a toxin containing seven disulfide bonds: bothropstoxin-1 from Bothrops jararacussu
2011 - BALDUINO, KELI N.; SPENCER, PATRICK J.; MALAVASI, NATALIA V.; CHURA-CHAMBI, ROSA M.; LEMKE, LAURA S.; MORGANTI, LIGIA