LC-MS and NMR characterization of the purple chromophore formed in the o-aminobenzaldehyde assay of dihydrodipicolinate synthase

V Mitsakos; SRA Devenish; PA O'Donnell; JA Gerrard; CA Hutton

Journal article

LC-MS and NMR characterization of the purple chromophore formed in the o-aminobenzaldehyde assay of dihydrodipicolinate synthase

V Mitsakos, SRA Devenish, PA O'Donnell, JA Gerrard, CA Hutton

Bioorganic and Medicinal Chemistry | PERGAMON-ELSEVIER SCIENCE LTD | Published : 2011

DOI: 10.1016/j.bmc.2010.12.029

Abstract

The enzyme dihydrodipicolinate synthase (DHDPS) has been widely investigated as a target for new antibiotics. The o-aminobenzaldehyde (o-ABA) assay is routinely used as a highly specific, if qualitative, tool for DHDPS purification, whereby fractions containing active DHDPS appear purple upon addition of o-ABA. The purple adduct absorbs in the visible region (540 nm) but has never been characterized in the 50 years since it was first reported. Structural characterization of this purple compound has been performed by UV spectrophotometry, NMR spectroscopy and tandem mass spectrometry. The extinction coefficient of this chromophore was also determined. © 2011 Elsevier Ltd. All rights reserved.

University of Melbourne Researchers

Craig Hutton Author

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Funding Acknowledgements

This work was supported in part by the Australian Research Council (DP0770888) and the Defense Threat Reduction Agency (Project ID AB07CBT004). SRAD was funded by the Foundation for Research, Science and Technology (contract UOCX0603). The authors acknowledge Tom Hennessy from Agilent Technologies for help with LC/ESI/TOF MS analysis.