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KMID : 0380020040190050394
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Korean Journal of Biotechnology and Bioengineering
2004 Volume.19 No. 5 p.394 ~ p.398
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Derivatization of α-Ketoacid for the Analysis of Aminotransferase Reaction Using Mass Spectrometry
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Lee Chang-Soo
Kim Eun-Gon
Kim Eun-Mi
Kim Byung-Ki
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Abstract
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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 -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
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KEYWORD
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Mass spectrometry, screening, derivatization, transaminase, substrate specificity
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