title: An advanced method for the determination of carboxyl methyl esterase activity using gas chromatography-chemical ionization-mass spectrometry.
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.
authors.affiliation: Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Yeon Jong
authors.initials: YJ
authors.lastname: Koo
authors.forename: Eunsil
authors.initials: E
authors.lastname: Yoon
authors.forename: Jong Tae
authors.initials: JT
authors.lastname: Song
authors.forename: Hak Soo
authors.initials: HS
authors.lastname: Seo
authors.forename: Jeong-Han
authors.initials: JH
authors.lastname: Kim
authors.forename: Yin-Won
authors.initials: YW
authors.lastname: Lee
authors.forename: Jong Seob
authors.initials: JS
authors.lastname: Lee
authors.forename: Jong-Joo
authors.initials: JJ
authors.lastname: Cheong
authors.forename: Yang Do
authors.initials: YD
authors.lastname: Choi
mesh_terms: D017360:Arabidopsis / Q000737:chemistry; D002265:Carboxylic Ester Hydrolases / Q000032:analysis*; D002855:Chromatography, Thin Layer; D004926:Escherichia coli / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007700:Kinetics; D010937:Plant Growth Regulators / Q000032:analysis; D010944:Plants / Q000737:chemistry; D012015:Reference Standards; D014297:Trimethylsilyl Compounds
chemical_list: D007202:Indicators and Reagents; D010937:Plant Growth Regulators; D014297:Trimethylsilyl Compounds; C047270:N,N-bis(trimethylsilyl)-2,2,2-trifluoroacetamide; C039293:trimethylchlorosilane; D002265:Carboxylic Ester Hydrolases