An advanced method for the determination of carboxyl methyl esterase activity using gas chromatography–chemical ionization–mass spectrometry
详细信息 查看全文
- 作者：Yeon Jong Koo ; Eunsil Yoon ; Jong Tae Song ; Hak Soo Seo ; Jeong-Han Kim ; Yin-Won Lee ; Jong Seob Lee ; Jong-Joo Cheong ; Yang Do Choi
- 关键词：Jasmonic acid methyl ester ; Salicylic acid methyl ester ; Indole 3-acetic acid methyl ester ; Carboxyl methyl esterase ; Gas chromatography– ; mass spectroscopy
- 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
- 出版年：2008
- 出版时间：15 February 2008
- 年：2008
- 卷：863
- 期：1
- 页码：80-87
- 全文大小：977 K
文摘
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.