title: Escherichia coli-Based Expression and In Vitro Activity Assay of 1-Aminocyclopropane-1-Carboxylate (ACC) Synthase and ACC Oxidase.
abstract: 1-Aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase are key enzymes in the ethylene biosynthetic pathway in plant tissues, and in vitro assay of their activities is indispensable for analysis, especially, for studying the action mechanism of inhibitors of ethylene biosynthesis. The enzymes can be obtained from plant tissues that are producing ethylene abundantly, such as ripening fruit- and senescing flower tissues, but it is necessary to separate the enzymes from co-extracted ACC by partial purification, making the procedure laborious and time-consuming. Here, we describe the production of the enzymes in Escherichia coli cells from corresponding cDNAs, and the procedures for assay of activities of the enzymes.
authors.affiliation: Faculty of Agriculture, Ryukoku University, Otsu, 520-2194, Japan. ssatoh@agr.ryukoku.ac.jp.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Satoh
authors.affiliation: Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Kosugi
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D004789:Enzyme Activation; D057075:Enzyme Assays / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D005030:Ethylenes / Q000096:biosynthesis; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D008190:Lyases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins
chemical_list: D005030:Ethylenes; D011994:Recombinant Proteins; C036216:ethylene; D000594:Amino Acid Oxidoreductases; D008190:Lyases; C021773:1-aminocyclopropanecarboxylate synthase
keywords: 1-Aminocyclopropane-1-carboxylate (ACC) synthase; ACC oxidase; Enzyme assay; Escherichia coli; Ethylene