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Separation and purification of penicillin acylase from Escherichia coli using AOT reverse micelles

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Biotechnology Techniques

Summary

The extraction of penicillin acylase by reverse micellar solutions of a surfactant was studied. A 50 mM solution of dioctyl sodium sulphosuccinate in isooctane extracted 46% of the enzyme activity in a crude periplasmic extract of induced cells of E. coli ATCC 9637. The increase in the specific activity of the final enzyme preparation, after stripping of the organic phase at pH 7.5, in the presence of 1 M KCl, was 8 - fold.

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Abbreviations

PA:

penicillin acylase (penicillin amidohydrolase EC 3.5.1.11)

AOT:

Aerosol OT (dioctyl sodium sulphosuccinate)

NIPAB:

6-nitro-3-(phenylacetamido)-benzoic acid

NABA:

6-nitro-3-aminobenzoic acid

BSA:

bovine serum albumin

SDS:

sodium dodecylsulphate

References

  • Aires-Barros, M. R. and Cabral, J. M. S. (1991) Biotechnol. Bioeng., 38, 1302 - 1307.

    Google Scholar 

  • Bruno, P., Caselli, M., Luisi, P. L., Maestro, M. and Traini, A. (1990). J. Phys. Chem., 94, 5908 - 5917.

    Google Scholar 

  • Cole, M., Savidge, T. and Vanderhaeghe, H. (1975). Methods in Enzymology, 43, 698–705.

    Google Scholar 

  • Dekker, M., Van't Riet, K. and Weijers, S.R. (1986). Chem. Eng., 33, B27-B33.

    Google Scholar 

  • Göklen, K. E. and Hatton, T. A. (1985). Biotechnol. Prog., 1, 69–74

    Google Scholar 

  • Kutzbach, C. and Rauenbusch, E. (1974). Hoppe-Seyler's Z. Physiol. Chem., 354, 45 - 53.

    Google Scholar 

  • Leser, M. E. and Luisi, P. L. (1989). Biotechnol. Tech., 3, 149 - 154.

    Google Scholar 

  • Leser, M. E. and Luisi, P. L. (1990). Chimia., 44, 270 - 282.

    Google Scholar 

  • Leser, M. E., Wei, G., Luisi, P. L. and Maestro, M. (1986). Biochem. Biophys.Res. Comm., 135, 629 - 635.

    Google Scholar 

  • Marcozzi, G., Correa, N., Luisi, P. L. and Caselli, M. (1991). Biotechnol. Bioeng., 38, 1239 - 1246.

    Google Scholar 

  • Rahaman, R. S., Chee, J. Y., Cabral, J. M. S. and Hatton, T. A. (1988). Biotechnol. Prog., 4, 218 - 224.

    Google Scholar 

  • Rodríguez, M., Güereca, L., Valle, F., Quintero, R. & López-Munguía, A. (1992). Process Biochem., 27, 217 - 223.

    Google Scholar 

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Authors and Affiliations

  1. Departamento de Química da Universidade do Minho, Campus de Gualtar, 4700, Braga, Portugal

    J. R. S. Alves & M. T. Ramalho

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

    L. P. Fonseca & J. M. S. Cabral

Authors
  1. J. R. S. Alves
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  2. L. P. Fonseca
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  3. M. T. Ramalho
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  4. J. M. S. Cabral
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Alves, J.R.S., Fonseca, L.P., Ramalho, M.T. et al. Separation and purification of penicillin acylase from Escherichia coli using AOT reverse micelles. Biotechnol Tech 9, 265–270 (1995). https://doi.org/10.1007/BF00151573

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  • DOI: https://doi.org/10.1007/BF00151573

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