Partial purification of a thylakoid-bound enzyme using temperature-induced phase partitioning.
Triton X-114 was used to partially purify broad bean polyphenol oxidase, a thylakoid membrane-bound enzyme, in latent form, free of phenolic compounds and chlorophylls, with a high recovery rate. The activation of the latent enzyme by detergents or trypsin was 10 times higher than that obtained when the enzyme was purified by other methods used in plant biochemistry, such as acetone powders and ammonium sulfate fractionation. The kinetic parameters of the latent and activated enzyme are also given.
Sánchez-Ferrer
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A
Departamento de Bioquímica, Facultad de Biología, Universidad de Murcia, Spain.
Bru
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R
García-Carmona
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F
D004156:Catechol Oxidase / Q000302:isolation & purification* / Q000378:metabolism; D002498:Centrifugation; D002736:Chloroplasts / Q000201:enzymology*; D004789:Enzyme Activation; D007887:Fabaceae; D007425:Intracellular Membranes / Q000201:enzymology; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D017830:Octoxynol; D010946:Plants, Medicinal; D011092:Polyethylene Glycols; D013696:Temperature
D008565:Membrane Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; D004156:Catechol Oxidase