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J Chromatogr B Analyt Technol Biomed Life Sci ; 1100-1101: 65-75, 2018 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-30292951

RESUMO

Dye-ligand affinity chromatography in a stirred fluidized bed has been developed for the rapid recovery of malate dehydrogenase (MDH) from highly turbid baker's yeast cell homogenate in a single step. The most suitable dye, namely Reactive Orange 4, in its optimal immobilized concentration of 8.78 mg/mL was immobilized onto high-density STREAMLINE matrix. To further examine optimal adsorption and elution conditions, the enzyme recovery operation was carried out using unclarified cell homogenates in stirred fluidized bed system. Aiming to develop a non-specific eluent, namely NaCl, to effectively elute the MDH adsorbed, direct recovery of MDH from highly turbid cell homogenate (50% w/v) in a stirred fluidized bed adsorption system was performed. The proposed system successfully achieved a recovery yield of 73.6% and a purification factor of 73.5 in a single step by using 0.6 M NaCl as an eluent at a high liquid velocity of 200 cm/h.


Assuntos
Extratos Celulares/química , Cromatografia de Afinidade/métodos , Malato Desidrogenase/isolamento & purificação , Saccharomyces cerevisiae/enzimologia , Adsorção , Compostos Azo/química , Corantes/química , Desenho de Equipamento , Concentração de Íons de Hidrogênio , Malato Desidrogenase/análise , Malato Desidrogenase/metabolismo , Ésteres do Ácido Sulfúrico/química
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