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Removal of proteases from Clostridium perfringens fermented broth by aqueous two-phase systems (PEG/citrate)

  • Original Paper
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Journal of Industrial Microbiology & Biotechnology

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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Acknowledgments

The authors wish to acknowledge the financial support of FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo, São Paulo State, Brazil), CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico, Brasilia, Brazil) and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Brasilia, Brazil).

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Authors and Affiliations

  1. Department of Biochemical and Pharmaceutical Technology, University of São Paulo, 05508-000, São Paulo, SP, Brazil

    Tatiana Souza Porto & Adalberto Pessoa Jr

  2. Department of Chemical Engineering, University of São Paulo, 05508-000, São Paulo, SP, Brazil

    Pedro Alcântara Pessôa-Filho

  3. Department of Fundamental Chemistry, UFPE, 50740-540, Recife, PE, Brazil

    Benício Barros Neto

  4. Laboratory of Immunopathology Keizo Asami (LIKA), UFPE, 50670-901, Recife, PE, Brazil

    José Luiz Lima Filho & Ana Lúcia Figueiredo Porto

  5. Department of Chemical and Process Engineering “G. B. Bonino”, via Opera Pia 15, 16145, Genoa, Italy

    Attilio Converti

Authors
  1. Tatiana Souza Porto
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  2. Pedro Alcântara Pessôa-Filho
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  3. Benício Barros Neto
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  4. José Luiz Lima Filho
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  5. Attilio Converti
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  6. Ana Lúcia Figueiredo Porto
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  7. Adalberto Pessoa Jr
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Correspondence to Tatiana Souza Porto.

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Porto, T.S., Pessôa-Filho, P.A., Neto, B.B. et al. Removal of proteases from Clostridium perfringens fermented broth by aqueous two-phase systems (PEG/citrate). J Ind Microbiol Biotechnol 34, 547–552 (2007). https://doi.org/10.1007/s10295-007-0230-8

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  • DOI: https://doi.org/10.1007/s10295-007-0230-8

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