Abstract
Penicillin G acylase (PGA) is used for the commercial production of semi-synthetic penicillins. It hydrolyses the amide bond in penicillin producing 6-aminopenicillanic acid and phenylacetate. 6-Aminopenicillanic acid, having the β-lactam nucleus, is the parent compound for all semi-synthetic penicillins. Penicillin G acylase from Kluyvera citrophila was purified and chemically modified to identify the role of arginine in catalysis. Modification with 20 mm phenylglyoxal and 50 mm 2,3-butanedione resulted in 82% and 78% inactivation, respectively. Inactivation was prevented by protection with benzylpenicillin or phenylacetate at 50 mm. The reaction followed psuedo-first order kinetics and the inactivation kinetics (V max, K m, and k cat) of native and modified enzyme indicates the essentiality of arginyl residue in catalysis.
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Kumar, R.S., Suresh, C., Pundle, A. et al. Evidence for the involvement of arginyl residue at the active site of penicillin G acylase from Kluyvera citrophila . Biotechnology Letters 26, 1601–1606 (2004). https://doi.org/10.1023/B:BILE.0000045660.65728.ba
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DOI: https://doi.org/10.1023/B:BILE.0000045660.65728.ba