Abstract
The synthesis of 2-nitro-5-[(hexanoyl)-amino]-benzoic acid and 2-nitro-5-[(octanoyl)-amino]-benzoic acid as chromogenic substrates for the determination of aliphatic penicillin acylase activity is described. During enzymatic hydrolysis, the released chromophore, 2-nitro-5-amino-benzoic acid, was detected at 405 nm. Penicillin acylase from Streptomyces lavendulae had an appreciable activity towards these substrates, which can then be used to detect penicillin acylases able to cleave hexanoyl and octanoyl residues off synthetic amides as well as penicillin dihydroF and penicillin K, their natural analogues.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
AlkemaWBL, Floris R, Janssen DB (1999) The use of chromogenic reference substrates for the kinetic analysis of penicillin acylases. Anal. Biochem. 275: 47-53.
Arroyo M, Torres R, de la Mata I, Castillón MP, Acebal C (2000) Prediction of penicillin V acylase stability in water-organic cosolvent monophasic systems as a function of solvent composition. Enzyme Microb. Technol. 27:122-126.
Baker WL (1979) Estimation of 6-aminopenicillanic acid by the ninhydrin reaction. Australian J. Pharm. Sci. 8: 86-90.
Baker WL (1983) Application of the fluorescamine reaction with 6-aminopenicillanic acid to estimation and detection of penicillin acylase activity. Antimicrob. Ag. Chemother. 23: 26-30.
Kerr DE (1993) A colorimetric assay for penicillin V amidase. Anal. Biochem. 209: 332-334.
Shewale JG, Kumar KK, Ambedkar GR (1987) Evaluation of determination of 6-amino penicillanic acid by p-dimethylaminobenzaldehyde. Biotechnol. Tech. 1: 69-72.
Torres R, de la Mata I, Castillón MP, Arroyo M, Torres J, Acebal C (1998) Purification and characterization of penicillin V acylase from Streptomyces lavendulae. In: Ballesteros A, Plou FJ, Iborra JL, Halling PJ, eds. Stability and Stabilization of Biocatalysts. Elsevier Science B.V.201-206.
Zhang Q, Zhang L, Han H, Zhang Y (1986) A method for screening penicillin G acylase-producing bacteria by means of 2-nitro-5-phenylacetaminobenzoic acid test paper. Anal. Biochem. 156: 413-416.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Arroyo, M., Torres-Guzmán, R., Torres-Bacete, J. et al. Chromogenic analogues of penicillin dihydroF and penicillin K for the continuous spectrophotometric determination of aliphatic penicillin acylase activity. Biotechnology Letters 24, 1045–1048 (2002). https://doi.org/10.1023/A:1016096111453
Issue Date:
DOI: https://doi.org/10.1023/A:1016096111453