Application of the fluorescamine reaction with 6-aminopenicillanic acid to estimation and detection of penicillin acylase activity.
نویسنده
چکیده
6-Aminopenicillanic acid may be quantitatively estimated by its reaction with fluorescamine in the concentration range of 1 to 10 micrograms/ml. The difference in reactivity between 6-aminopenicillanic acid and benzylpenicillin, which does not react with fluorescamine, can be used to determine penicillin acylase activity and obtain data on enzyme parameters and inhibitors. Unlike amino acids and peptides, 6-aminopenicillanic acid reacts strongly with fluorescamine at pH 4, an observation which can be used to determine the presence of penicillin acylase in whole bacterial cell preparations.
منابع مشابه
Isolation of a Penicillin Acylase Producing E.coli and Kinetic Characterization of the Whole Cell Enzyme Activity
Penicillin acylase (EC 3.5.1.11) has been a target of study for a long time because of its pivotal role in the deacylation of the penicillin into the 6- aminopenicillanic acid (6-APA) and the side-chain organic acids. This property of penicillin acylase has been exploited commercially for large scale production of 6-APA, which is the key intermediate in the manufacture of semi-synthetic penicil...
متن کاملIsolation of a Penicillin Acylase Producing E.coli and Kinetic Characterization of the Whole Cell Enzyme Activity
Penicillin acylase (EC 3.5.1.11) has been a target of study for a long time because of its pivotal role in the deacylation of the penicillin into the 6aminopenicillanic acid (6-APA) and the side-chain organic acids. This property of penicillin acylase has been exploited commercially for large scale production of 6-APA, which is the key intermediate in the manufacture of semi-synthetic penicilli...
متن کاملOptimization of Enzymatic Synthesis of Ampicillin Using Cross-Linked Aggregates of Penicillin G Acylase
Penicillin G acylase from E. coli TA1 was immobilized by Cross-Linked Enzyme Aggregates (CLEA), a new method for immobilization. This biocatalyst and commercial immobilized penicillin G acylase (PGA-450) were used to study the effect of pH, temperature and substrate concentration on the synthesis of ampicillin from phenyl glycine methyl ester (PGME) and 6-aminopenicillanic acid (6-APA). Compare...
متن کاملOptimization of Enzymatic Synthesis of Ampicillin Using Cross-Linked Aggregates of Penicillin G Acylase
Penicillin G acylase from E. coli TA1 was immobilized by Cross-Linked Enzyme Aggregates (CLEA), a new method for immobilization. This biocatalyst and commercial immobilized penicillin G acylase (PGA-450) were used to study the effect of pH, temperature and substrate concentration on the synthesis of ampicillin from phenyl glycine methyl ester (PGME) and 6-aminopenicillanic acid (6-APA). Compare...
متن کاملPenicillin acylase from the hybrid strains Escherichia coli 5K(pHM12): enzyme formation and hydrolysis of beta-lactam antibiotics with whole cells.
Penicillin acylase formation by the hybrid strain Escherichia coli 5K(pHM12) was studied under different culture conditions and reached 200 to 250 mumol of 6-aminopenicillanic acid per min per g of bacteria (wet weight) for penicillin G. The Km of whole-cell acylase was determined with 9 to 11 mM for penicillin G at a pH optimum of 7.8 at 45 degrees C. A competitive product inhibition for pheny...
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ورودعنوان ژورنال:
- Antimicrobial agents and chemotherapy
دوره 23 1 شماره
صفحات -
تاریخ انتشار 1983