Hielscher Björn, Charton Lennart, Mettler-Altmann Tabea, Linka Nicole
Institute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, Universitätsstrasse 1, Düsseldorf, 40225, Germany.
Methods Mol Biol. 2017;1595:291-304. doi: 10.1007/978-1-4939-6937-1_27.
Peroxisomal β-oxidation in plants is essential for mobilization of storage oil in seed-oil storing plants, such as Arabidopsis thaliana. In plants, degradation of fatty acids occurs exclusively in peroxisomes via β-oxidation, driving seedling growth and development upon germination. Thus, the determination of storage oil breakdown rates is a useful approach to investigate defects in peroxisomal β-oxidation. Here we describe an acid catalyzed derivatization process of fatty acids representing a fast and efficient procedure to generate high yields of fatty acid methyl esters (FAMEs). The subsequent analysis by gas chromatography coupled to mass spectrometry (GC-MS) allows the quantification of total fatty acid content. The results provide detailed information of the complete storage oil breakdown process via peroxisomal β-oxidation during seedling growth.
过氧化物酶体β-氧化在植物中对于种子储存油的植物(如拟南芥)中储存油的动员至关重要。在植物中,脂肪酸的降解仅通过β-氧化在过氧化物酶体中发生,驱动种子萌发时的幼苗生长和发育。因此,测定储存油的分解速率是研究过氧化物酶体β-氧化缺陷的一种有用方法。在这里,我们描述了一种脂肪酸的酸催化衍生化过程,该过程是一种快速有效的方法,可高产率地生成脂肪酸甲酯(FAMEs)。随后通过气相色谱-质谱联用(GC-MS)进行分析,可以对总脂肪酸含量进行定量。结果提供了幼苗生长过程中通过过氧化物酶体β-氧化进行的完整储存油分解过程的详细信息。