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Derivatization of a-ketoacid for the Analysis of Aminotransferase Reaction Using Mass Spectrometry  

Lee Chang Soo (Department of Chemical Engineering, Chungnam National University)
Kim Yun Gon (School of Chemical Engineering and Institute of Molecular Biology and Genetics, Seoul National University)
Kim Eun Mi (School of Chemical Engineering and Institute of Molecular Biology and Genetics, Seoul National University)
Kim Byung Gee (School of Chemical Engineering and Institute of Molecular Biology and Genetics, Seoul National University)
Publication Information
KSBB Journal / v.19, no.5, 2004 , pp. 394-398 More about this Journal
Abstract
A derivatization method in mass spectrometry for small molecular analysis was developed to solve the problems of volatility of many analytes, difficult ionization of analytes, and undiscriminating isobaric analytes. This derivatization method, oximation of $\alpha$-ketoacid, in the transaminase reaction leads to change of mass difference between amine reactants and ketoacid products. In addition, regardless of the kinds of ketoacid, the linear relationship between the peak intensity and its concentration (from 1 mM to 10 mM) shows that quantitative analysis of the conversion of the reaction can be executed by the analysis of peak intensity of the corresponding oximated ketoacids. Furthermore, this method can be used for identifying transaminase as well as determining its substrate specificity
Keywords
Mass spectrometry; screening; derivatization; transaminase; substrate specificity;
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