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Pure Appl. Chem. 76(4), 765-775, 2004

Pure and Applied Chemistry

Vol. 76, Issue 4

Glutamate oxidase advances in the selective bioanalytical detection of the neurotoxic amino acid b-ODAP in grass pea: A decade of progress

G. Moges, N. Wodajo, L. Gorton, Y. Yigzaw, K. Kalcher, A. Belay, G. Akalu, B. M. Nair, and T. Solomon

Department of Chemistry, Addis Ababa, Ethiopia; Department of Analytical Chemistry, Institute of Chemistry, Karl-Franzens University, Graz, Austria; Department of Analytical Chemistry, University of Lund, Sweden; Applied Nutrition and Food Chemistry, University of Lund, Sweden

Abstract: The search for an enzyme as a reagent for selective bioanalytical detection of the neurotoxic amino acid, b-N-oxalyl L-alpha, b-diaminopropionic acid, b-ODAP (found in grass pea, Lathyrus sativus) led to its redox catalytic reaction by glutamate oxidase (GluOx). Homogeneous kinetic studies and an immobilized GluOx reactor-based flow-injection assay were initially made for beta-ODAP with small immobilized GluOx/catalase glutamate destroying prereactors. The method was applied to examine the toxin content in processed grass pea. The kinetics and the equilibrium of the thermal isomerization of b-ODAP to the nontoxic isomer a-ODAP established that GluOx is specific to the neurotoxin. The first ever GluOx-based amperometric biosensor for liquid chromatography (LC) detection was reported in 1997. This biosensor coupled with a refractive index detector improved LC performance. The most recent work with GluOx resulted in MnO2-based screen-printed amperometric biosensor, with offline elimination of glutamate interference by glutamate decarboxylase. A single-shot chemiluminescent sensor developed for hydrogen peroxide is also proposed for b-ODAP with GluOx application. This decade of progress resulted from studies that included four Ph.D. (Ethiopia, Sweden, Austria), four M.Sc. (Ethiopia, Sweden) and Licentiate (Sweden) theses projects, plus one collaborative project in Sweden. The advances in grass pea research may be regarded as a model north–south cooperation for research and education.

*Plenary lectures presented at the Inaugural Conference for the Southern and Eastern Africa Network of Analytical Chemists (SEANAC), Gaborone, Botswana, 7-10 July 2003. Other presentations are published in this issue, pp. 697-888.

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