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Studies on the extraction and back-extraction of a recombinant cutinase in a reversed micellar extraction process

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Abstract

This work deals with the extraction and back-extraction of a recombinant cutinase using AOT reversed micelles in isooctane. The effect of pH, ionic strength, AOT concentration and temperature on the extraction and back-extraction of the cutinase was investigated. High extraction (97%) of the cutinase was achieved at pH 7.0 with a 50 mM Tris-HCl buffer solution containing 100 mM KCl, but a low activity was detected in the reversed micellar phase. At pH 9.0, cutinase was extracted (75%) to the reversed micelles with higher activity. Cutinase was recovered (50%) from a reversed micellar phase (100 mM AOT/isooctane) into a 50 mM Tris-HCl buffered solution at pH 9.0 with 100 mM KCl, and 20°C. Protein and cutinase activity global yields of 38 and 45%, respectively, were obtained for the global process, extraction and back-extraction steps, using low ionic strength, pH 9.0, 100 mM AOT and 20°C.

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Authors and Affiliations

  1. Instituto Tecnológico do Estado de Pernambuco-ITEP Recife, Pernambuco, Brasil

    M. G. Carneiro-da-Cunha

  2. Laboratório de Engenharia Bioquímica, Instituto Superior Técnico, 1000, Lisboa, Portugal

    J. M. S. Cabral & M. R. Aires-Barros

Authors
  1. M. G. Carneiro-da-Cunha
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  2. J. M. S. Cabral
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  3. M. R. Aires-Barros
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Additional information

Maria das Graças Carneiro da Cunha acknowledges a Ph.D. fellowship from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Centro de Pesquisas Aggeu Magalhães, Brasil. This work was partly financed by the BRIDGE Programme (Contract BIOT-CT91-0274(DTEE)).

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Carneiro-da-Cunha, M.G., Cabral, J.M.S. & Aires-Barros, M.R. Studies on the extraction and back-extraction of a recombinant cutinase in a reversed micellar extraction process. Bioprocess Engineering 11, 203–208 (1994). https://doi.org/10.1007/BF00369631

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