Production of 6-aminopenicillanic acid in aqueous two-phase systems by recombinant Escherichia coli with intracellular penicillin acylase |
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Authors: | Cao Xue-jun Wu Xing-yan Luis J.P. Fonseca Joaquim M.S. Cabral João Carlos Marcos |
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Affiliation: | State Key Laboratory of Bioreactor Engineering, Department of Biochemical Engineering, East China University of Science and Technology, Shanghai 200237, PR China. |
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Abstract: | Bioconversion of penicillin G in PEG 20000/dextran T 70 aqueous two-phase systems was achieved using the recombinant Escherichia coli A56 (ppA22) with an intracellular penicillin acylase as catalyst. The best conversion conditions were attained for: 7% (w/v) substrate (penicillin G), enzyme activity in bottom phase 52 U ml(-1), pH 7.8, temperature 37 degrees C, reaction time 40 min. Five repeated batches could be performed in these conditions. Conversions ratios between 0.9-0.99 mol of 6-aminopenicillanic acid (6-APA) per mol of penicillin G, were obtained and volumetric productivity was 3.6-4.6 micromol min(-1) ml(-1). In addition the product 6-APA could be directly crystallized from the top phase with a purity of 96%. |
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Keywords: | 6-aminopenicillanic acid production aqueous two-phase systems bioconversion penicillin acylase recombinant E. coli A56 |
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