Production, purification, characterization and immunological studies of E.coli E-U₁₀ penicillin amidase

Temsah, S.

Production, purification, characterization and immunological studies of E.coli E-U₁₀ penicillin amidase [electronic reource]. - p.1-16.

Includes references.

Penicillin amidase enzyme (PA) was isolated from the culture filtrate of one isolated strain of E. coli E-U₁₀ grown on optimized medium after 24 hours incubation. The enzyme was purified to homogeneity by sequential ammonium sulfate fractionation and ion exchange and gel filtration chromatography. On using Disc-PAGE, the final purification fraction showed only one distinctive band indicating high purity. On using SDS-PAGE the final fraction showed two distinctive units with molecular weight of 22000 and 59000 Dalton. The optimal pH, temperature and time course for activity were 7.5, 37°C and 10 minutes respectively. The enzyme with penicillin G as substrate had a Km of 13 mM and Vmax 3.92 U/ml.


Escherichia coli.

penicillin amidase.

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