Affinity chromatography of an ethylene-synthesizing enzyme from red algaPorphyra teneraon an immobilized inhibitor of ethylene evolution

摘要

A method was established to purify acrylate decarboxylase fromPorphyra teneraby affinity chromatography using a proteinaceous inhibitor of ethylene evolution in marine algae, isolated fromP. teneraas a ligand. The proteinaceous inhibitor was covalently coupled to porous glass via four different spacer arms. The porous glass-aminopropyltriethoxysilane-succinatephenylendiamine-succinate-inhibitor appeared to be the best derivative for retaining acrylate decarboxylase extracted fromP. tenera.Acrylate decarboxylase was extracted from 10 kg ofP. teneraand semi-purified by ammonium sulfate. preparation and gel filtration on Sephadex G-100. The active fraction was applied to an affinity column. Acrylate decarboxylase was eluted in the starting buffer containing 0.2mNaCl. Ethylene formation from acrylate was detected in the presence of this enzyme extract, but not in the case of the boiled enzyme extract. Acrylate decarboxylase was inhibited by the inhibitor isolated fromP. tenera. These facts indicate that the formation of ethylene in marine algae from acrylate proceeds enzymatically.