Resolution of DL-Phenylglycine by Penicillin G acylase.

The parameters for complete hydrolysis of L-phenyl acetyl phenylglycine (L-PAPG) using immobilized penicillin G acylase (IMEPGA) were investigated. IMEPGA exhibited maximum activity at pH 8.5 and 50 degrees C. The apparent Km value observed was 10 mM. Quantitative hydrolysis (>97%) of the L-PAPG was achieved within 45 min, at pH 7.8 and 37 degrees C, when 0.5% (w/v) of DL-PAPG was used and the concentration of IMEPGA was 133 IU/gm of DL-PAPG. The IMEPGA was used for 50 cycles.

Isolation and purification of penicillin G acylase obtained from Escherichia coli (NCIM-2400) and immobilisation on Eupergit C for the production of 6 amino penicillanic acid.

Penicillin G-Acylase is produced by submerged cultivation of E. Coli (NCIM-2400) and extracted from the harvested fermented broth, purified (affinity chromatography) and immobilised on Eupergit C (Synthetic polymer in bead form). The immobilised penicillin G acylase properties are studied and compared with soluble penicillin G-acylase. The control parameters for conversion of penicillin G-K to 6 APA are optimised [e.g. substrate (Pen G-K) concentration ratio to immobilised penicillin G-acylase, temperature, pH etc.] in a stirred tank reactor. Our findings suggest that immobilised penicillin G-acylase can be used commercially and the productivity of 1 kg. of immobilised enzyme is around 400 kg of 6 APA under given desired stipulated conditions.