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1.
Biotechnol J ; 7(5): 649-55, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-21905227

RESUMO

Urea has been considered as a promising alternative nitrogen source for the cultivation of Arthrospira platensis if it is possible to avoid ammonia toxicity; however, this procedure can lead to periods of nitrogen shortage. This study shows that the addition of potassium nitrate, which acts as a nitrogen reservoir, to cultivations carried out with urea in a fed-batch process can increase the maximum cell concentration (X(m) ) and also cell productivity (P(X) ). Using response surface methodology, the model indicates that the estimated optimum X(m) can be achieved with 17.3 mM potassium nitrate and 8.9 mM urea. Under this condition an X(m) of 6077 ± 199 mg/L and a P(X) of 341.5 ± 19.1 mg L(-1) day(-1) were obtained.


Assuntos
Biotecnologia/métodos , Nitratos/metabolismo , Compostos de Potássio/metabolismo , Spirulina/crescimento & desenvolvimento , Spirulina/metabolismo , Ureia/metabolismo , Biomassa , Técnicas de Cultura de Células , Proliferação de Células , Meios de Cultura/química , Meios de Cultura/metabolismo , Concentração de Íons de Hidrogênio , Análise Multivariada , Nitrogênio/metabolismo , Análise de Regressão
2.
Biotechnol Prog ; 26(6): 1644-53, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20572295

RESUMO

In biotechnology, endotoxin (LPS) removal from recombinant proteins is a critical and challenging step in the preparation of injectable therapeutics, as endotoxin is a natural component of bacterial expression systems widely used to manufacture therapeutic proteins. The viability of large-scale industrial production of recombinant biomolecules of pharmaceutical interest significantly depends on the separation and purification techniques used. The aim of this work was to evaluate the use of aqueous two-phase micellar system (ATPMS) for endotoxin removal from preparations containing recombinant proteins of pharmaceutical interest, such as green fluorescent protein (GFPuv). Partition assays were carried out initially using pure LPS, and afterwards in the presence of E. coli cell lysate. The ATPMS technology proved to be effective in GFPuv recovery, preferentially into the micelle-poor phase (K(GFPuv) < 1.00), and LPS removal into the micelle-rich phase (%REM(LPS) > 98.00%). Therefore, this system can be exploited as the first step for purification in biotechnology processes for removal of higher LPS concentrations.


Assuntos
Biotecnologia , Fracionamento Químico/métodos , Escherichia coli/metabolismo , Fermentação , Lipopolissacarídeos/isolamento & purificação , Escherichia coli/química , Escherichia coli/citologia , Proteínas de Fluorescência Verde/química , Lipopolissacarídeos/química , Lipopolissacarídeos/metabolismo , Micelas , Água/química
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