Enzymatic process for the determination of beta-lactam antibioti

Details Enzymatic process for the determination of beta-lactam antibioti Enzymatic process for the determination of beta-lactam antibioti

1983-01-31

1985-10-08

Rosen, Sam

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving hydrolase

435180, C12Q 136

Patent

active

045460760

ABSTRACT:
An enzymatic process for the determination of beta-lactam antibiotics in a biological liquid, comprising the steps of: (1) incubating the liquid with soluble D-alanyl-D-alanine-carboxypeptidase produced by Actinomadura R 39, said enzyme being immobilized on a water-insoluble support, the beta-lactam antibiotic reacting with the immobilized enzyme to form an inactive and equimolecular enzyme-antibiotic complex; (2) separating the immobilized enzyme from the liquid and washing it; (3) incubating the immobilized enzyme with a substrate solution to hydrolyze it and form an amount of D-alanine corresponding to the residual enzymatic activity; (4) determining the amount of D-alanine formed; and (5) comparing the determination of step (4) with a standard to obtain the concentration of the antibiotic in the biological liquid; as well as a test set for carrying out this process and comprising the necessary reagents.

REFERENCES:
patent: 4170696 (1979-10-01), Hirohara et al.
Fuad et al.--Biochem. J., vol. 155, (1976), pp. 623-629.
Frere et al., Antimicrobial Agents and Chemotherapy, vol. 18, No. 4, Oct. 1980, pp. 506-510.
Chemical Abstracts, vol. 90, No. 25, Jun. 18, 1979, p. 227, No. 199525M.
Chemical Abstracts, vol. 85, No. 23, Dec. 6, 1976, p. 180, No. 173261U.
Chemical Abstracts, vol. 87, No. 7, Aug. 15, 1977, p. 73, No. 48470b.
Chemical Abstracts, vol. 88, No. 5, Jan. 30, 1978, pp. 187-188, No. 33780Z.

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