1. An advanced method for the determination of carboxyl methyl esterase activity using gas chromatography–chemical ionization–mass spectrometry
- Author
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Jong Tae Song, Yin-Won Lee, Jong Seob Lee, Hak Soo Seo, Eunsil Yoon, Jeong-Han Kim, Yang Do Choi, Jong-Joo Cheong, and Yeon Jong Koo
- Subjects
Trimethylsilyl Compounds ,Silylation ,Esterase Gene ,Clinical Biochemistry ,Arabidopsis ,Biochemistry ,Esterase ,Gas Chromatography-Mass Spectrometry ,Analytical Chemistry ,chemistry.chemical_compound ,Plant Growth Regulators ,Escherichia coli ,Derivatization ,Chemical ionization ,Chromatography ,Cell Biology ,General Medicine ,Hydrogen-Ion Concentration ,Plants ,Reference Standards ,Kinetics ,chemistry ,Reagent ,Indicators and Reagents ,Chromatography, Thin Layer ,Gas chromatography ,Carboxylic Ester Hydrolases ,Salicylic acid - Abstract
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.
- Published
- 2008
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