Removal of proteases from <Emphasis Type="Italic">Clostridium perfringens</Emphasis> fermented broth by aqueous two-phase systems (PEG/citrate) |
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| Authors: | Tatiana Souza Porto Pedro Alcântara Pessôa-Filho Benício Barros Neto José Luiz Lima Filho Attilio Converti Ana Lúcia Figueiredo Porto Jr" target="_blank">Adalberto PessoaJr |
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| Institution: | 1.Department of Biochemical and Pharmaceutical Technology,University of S?o Paulo,S?o Paulo,Brazil;2.Department of Chemical Engineering,University of S?o Paulo,S?o Paulo,Brazil;3.Department of Fundamental Chemistry,UFPE,Recife,Brazil;4.Laboratory of Immunopathology Keizo Asami (LIKA), UFPE,Recife,Brazil;5.Department of Chemical and Process Engineering “G. B. Bonino”,Genoa,Italy |
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| Abstract: | In order to reduce the toxicity of Clostridium perfringens fermentation broths used in vaccine preparation, we developed two-phase aqueous systems for removal of toxin-activating proteases.
Removal of the proteases inhibits the conversion of protoxin to active toxin. In order to establish the conditions under which
the phase separation occurs, binodal curves, formed by poly(ethylene glycol) (PEG) and sodium citrate, were investigated at
different values of pH and PEG molar mass. A 24-experimental design was used to evaluate the influence of PEG molar mass and concentration, citrate concentration and pH
on protease partition coefficient, removal factor and protease removal yield. It has been found that simultaneous increase
in PEG molar mass and decrease in citrate concentration remarkably improved the removal factor, whereas the protease removal
yield showed an opposite trend. The best conditions for the system under consideration (removal factor of 2.69 and yield of
116%) were obtained at pH 8.0 using PEG molar mass of 8000 g mol−1 and concentrations of PEG and citrate of 24 and 15%, respectively. |
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| Keywords: | Extraction ATPS Experimental design C perfringens Protease Detoxification |
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