Glucose oxidase and catalase adsorption onto Cibacron Blue F3GA-attached microporous polyamide hollow-fibres

The aim of this study was to explore in detail the performance of polyamide hollow fibers to which Cibacron Blue F3GA was attached for adsorption of proteins. Model proteins were glucose oxidase, as a flavo-enzyme and contains two tightly bound flavine adenine dinucleotide cofactor, and catalase as a heme-containing metallo-enzyme. The hollow fiber structure was characterized by scanning electron microscopy. These dye-carrying hollow-fibers (35.8 mumol/g) were used in the glucose oxidase and catalase adsorption-elution studies. The non-specific adsorption values were 1.25 mg/g for glucose oxidase and 1.97 mg/g for catalase. Cibacron Blue F3GA attachment increased the adsorption capacity up to 248 mg/g. Langmuir adsorption model was found to be applicable in interpreting glucose oxidase and catalase adsorption by Cibacron Blue F3GA-attached hollow fibers. Significant amount of the adsorbed proteins (up to 97%) was eluted in I h in the elution medium containing 1.0 M NaSCN at pH 8.0. In order to determine the effects of adsorption and elution conditions on possible conformational changes of studied protein structures, fluorescence spectrophotometry was employed. It was concluded that polyamide dye-affinity hollow-fibers can be applied for glucose oxidase and catalase adsorption without causing any significant conformational changes. Repeated adsorption/elution processes showed that these dye-attached hollow-fibers are suitable for glucose oxidase and catalase separation. (C) 2002 Published by Elsevier Science B.V.