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Assay of penicillin acylase in organic media

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Biotechnology Techniques

Summary

The synthetic substrate 6-nitro-3-(phenylacetamido) benzoic acid (NIPAB) is an appropriate substrate for assaying penicillin acylase activity in reversed micellar systems of Aerosol - OT in isooctane. Accumulation of 6-nitro-3-aminobenzoic acid (NABA) produced by the enzymatic hydrolysis of NIPAB, followed by the increase in absorbance at 405 nm, was linear at 4 to 20 mM for up to 30 minutes and 15 °C to 40 °C.

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Abbreviations

PA:

penicillin acylase (penicillin amidohydrolase EC 3.5.1.11)

AOT:

Aerosol OT (sodium bis- (2-ethylhexyl) sulfosuccinate)

NIPAB:

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

NABA:

6-nitro-3-aminobenzoic acid

BSA:

bovine serum albumin

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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. Assay of penicillin acylase in organic media. Biotechnol Tech 9, 729–730 (1995). https://doi.org/10.1007/BF00159238

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  • DOI: https://doi.org/10.1007/BF00159238

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