Separation and purification of penicillin acylase from Escherichia coli using AOT reverse micelles |
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Authors: | J R S Alves L P Fonseca M T Ramalho J M S Cabral |
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Institution: | (1) Departamento de Química da Universidade do Minho, Campus de Gualtar, 4700 Braga, Portugal;(2) Laboratório de Engenharia Bioquímica, Instituto Superior Técnico, 1000 Lisboa, Portugal |
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Abstract: | 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.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 |
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