Open AccessPurification and partial characterization of 1‐aminocyclopropane‐1‐carboxylate synthase from tomato pericarp
Author(s) -
STRAETEN Dominique,
WIEMEERSCH Luc,
GOODMAN Howard M.,
MONTAGU Marc
Publication year - 1989
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1989.tb14873.x
Subject(s) - carboxylate , enzyme , chemistry , atp synthase , methionine , substrate (aquarium) , biochemistry , chromatography , gel electrophoresis , biology , amino acid , ecology
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 μg enzyme.