Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 863, Issue 1, Pages 80-87Publisher
ELSEVIER
DOI: 10.1016/j.jchromb.2008.01.015
Keywords
jasmonic acid methyl ester; salicylic acid methyl ester; indole 3-acetic acid methyl ester; carboxyl methyl esterase; gas chromatography-mass spectroscopy
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We developed a quantitative method for the determination of methyl esterase activity, analyzing substrate specificity against three major signal molecules, jasmonic acid methyl ester (MeJA), salicylic acid methyl ester (MeSA), and indole-3-acetic acid methyl ester (MeIAA). We used a silylation reagent for chemical derivatization and used gas chromatography (GC)-mass spectroscopy in analyses, for high precision. To test this method, an Arabidopsis esterase gene, AtME8, was expressed in Escherichia coli, and then the kinetic parameters of the recombinant enzyme were determined for three substrates. Finally, this method was also applied to the direct quantification of phytohormones in petals from lilies and roses. (C) 2008 Elsevier B.V. All fights reserved.
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