Extraction of Ascorbate Oxidase from <Emphasis Type="Italic">Cucurbita maxima</Emphasis> by Continuous Process in Perforated Rotating Disc Contactor Using Aqueous Two-Phase Systems |
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| Authors: | T S Porto P P Marques C S Porto K A Moreira J L Lima-Filho A Converti A Pessoa A L F Porto |
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| Institution: | 1.Departament of Biochemical and Pharmaceutical Technology,University of Sao Paulo,Sao Paulo,Brazil;2.Laboratory of Immunopathology Keizo Asami (LIKA),University Federal of Pernambuco,Recife,Brazil;3.Academic Unit of Garanhuns,University Federal Rural of Pernambuco,Recife,Brazil;4.Department of Chemical and Process Engineering,University of Genoa,Genoa,Italy;5.Department of Animal Morphology and Physiology,University Federal Rural of Pernambuco,Recife,Brazil |
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| Abstract: | The ascorbate oxidase is the enzyme used to determine the content of ascorbic acid in the pharmaceutical and food industries
and clinics analyses. The techniques currently used for the purification of this enzyme raise its production cost. Thus, the
development of alternative processes and with the potential to reduce costs is interesting. The application of aqueous two-phase
system is proposed as an alternative to purification because it enables good separation of biomolecules. The objective of
this study was to determine the conditions to continuously pre-purify the enzyme ascorbate oxidase by an aqueous two-phase
system (PEG/citrate) using rotating column provided with perforated discs. Under the best conditions (20,000 g/mol PEG molar
mass, 10% PEG concentration, and 25% citrate concentration), the system showed satisfactory results (partition coefficient,
3.35; separation efficiency, 54.98%; and purification factor, 1.46) and proved suitable for the pre-purification of ascorbate
oxidase in continuous process. |
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