title: Purification and partial characterization of 1-aminocyclopropane-1-carboxylate synthase from tomato pericarp.
abstract: 1-Aminocyclopropane-1-carboxylate synthase was purified 5000-fold from LiCl-induced tomato fruit slices by conventional and high-performance liquid chromatography. The final preparation was estimated to be between 25% and 50% pure. Two-dimensional gel electrophoresis indicates that 1-aminocyclopropane-1-carboxylate synthase activity is associated with a 45-kDa polypeptide, with a pI of 5.8 +/- 0.2. The enzyme is inactivated both by its substrate, S-adenosyl-L-methionine (AdoMet) and by one of its products, 1-aminocyclopropane-1-carboxylate. Due to the extremely low abundance of the protein it was necessary to scale up the extraction in order to obtain reasonable amounts for sequence analysis. Therefore, 200 kg tomatoes were extracted on semi-industrial scale and 1-aminocyclopropane-1-carboxylate synthase purified. This yielded approximately 150 micrograms enzyme.
authors.affiliation: Laboratorium voor Genetica, Rijksuniversiteit Gent, Belgium.
authors.forename: D
authors.initials: D
authors.lastname: Van der Straeten
authors.forename: L
authors.initials: L
authors.lastname: Van Wiemeersch
authors.forename: H M
authors.initials: HM
authors.lastname: Goodman
authors.forename: M
authors.initials: M
authors.lastname: Van Montagu
mesh_terms: D000595:Amino Acid Sequence; D002021:Buffers; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005638:Fruit / Q000201:enzymology*; D005990:Glycerol / Q000494:pharmacology; D006358:Hot Temperature; D007526:Isoelectric Point; D008190:Lyases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D008750:Methyldopa / Q000494:pharmacology
chemical_list: D002021:Buffers; D008750:Methyldopa; D008190:Lyases; C021773:1-aminocyclopropanecarboxylate synthase; D005990:Glycerol