Extraction penicillin G acylase from Alcaligenes faecalis in recombinant Escherichia coli with cetyl-trimethylammoniumbromide

Cells were permeabilized with cetyl-trimethylammoniumbromide (CTAB) to extract penicillin G acylase (sp. Alcaligenes faecalis) in recombinant Escherichia coli. The optimum conditions were 0.5% (w/v) CTAB, 0.5 M ionic strength, pH 8.0 and at 4 °C for 28 h. The maximum relative enzyme activity obtained was 93.4%, and the specific activity increased about 1.6 times than that by ultrasonic fragmentation. Furthermore, CTAB-extracted enzyme solution had fine stability, and almost no activity decreased as the enzyme was preserved at 4 °C for 30 days. From these results, the method can be used as a high-performance system about the extraction of penicillin G acylase from A. faecalis that resides mainly in the outer periplasmic space of E. coli.