Determination of Carboxypeptidase Activity in Clinical Pathogens by Gas Chromatography–Mass Spectrometry

Lough, Fraser, Perry, John, Stanforth, Stephen and Dean, John (2016) Determination of Carboxypeptidase Activity in Clinical Pathogens by Gas Chromatography–Mass Spectrometry. Analytical Letters, 49 (8). pp. 1272-1277. ISSN 0003-2719

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Official URL: http://dx.doi.org/10.1080/00032719.2015.1092153

Abstract

A novel method for the determination of benzoic acid has been employed to identify carboxypeptidase activities in clinically relevant pathogens. Benzoic acid was determined after chemical derivatization by gas chromatography–mass spectrometry (GC–MS). N-Benzoyl amino acid substrates were evaluated for the detection of carboxypeptidase activities in a number of clinical pathogens. Upon enzymatic hydrolysis of these substrates, benzoic acid was produced which was detected by extraction from the liquid culture supernatant, derivatization as the trimethylsilyl ester, with subsequent analysis by GC–MS. Enzymatic hydrolysis of N-benzoyl glycine was observed for S. agalactiae, M. morganii, and A. baumannii. In addition, P. fluorescens was found to hydrolyze N-benzoyl-L-glutamic acid. Although the method provides an alternative approach for determining carboxypeptidase activity, ultimately it would not be a suitable method in a clinical setting. However, the method is well-suited for identifying carboxypeptidase activities that have not been previously described or to corroborate a carboxypeptidase assay with the ninhydrin reagent.

Item Type: Article
Uncontrolled Keywords: Carboxypeptidase activity, chemical derivatization, enzymatic hydrolysis, gas chromatography–mass spectrometry: N-benzoyl amino acid substrates
Subjects: F100 Chemistry
Department: Faculties > Health and Life Sciences > School of Life Sciences > Applied Sciences
Depositing User: Becky Skoyles
Date Deposited: 17 May 2016 08:11
Last Modified: 14 May 2017 07:20
URI: http://nrl.northumbria.ac.uk/id/eprint/26798

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