Purification and properties of amine oxidase from pea seedlings
نویسندگان
چکیده
Amine oxidase (EC 1.4.3.6) was purified from pea (Pisum sativum) seedlings with the aim of characterizing its catalytic properties. The specific activity and selectivity of the enzyme were studied with an oxygen sensor by following the kinetics of the amine oxidation reaction, catalysed by the enzyme. The enzyme catalytic constants were calculated from the transient signal of the oxygen sensor with the help of a model proposed earlier for amperometric biosensors. The pea seedlings amine oxidase had the highest activity towards putrescine and cadaverine; a very low or zero activity was registered towards other studied amines. To characterize different steps of the purification process, we compared the activity of enzyme preparations towards 0.15 mM cadaverine. The molecular mass of the purified enzyme was 184.0 ± 2.6 kDa.
منابع مشابه
Purification and characterization of pea seedling amine oxidase for crystallization studies.
Pea (Pisum sativum L.) seedling amine oxidase (EC 1.4.3.6) is the first amine oxidase to be crystallized that diffracts to atomic resolution (2.5 A). Extensive modifications of a published purification procedure were necessary to obtain protein that would give diffraction-quality crystals. Here we report the improved purification and also use this high-purity protein to reexamine some fundament...
متن کاملPea (Pisum sativum) diamine oxidase contains pyrroloquinoline quinone as a cofactor.
Diamine oxidase was prepared from pea (Pisum sativum) seedlings by a new purification procedure involving two h.p.l.c. steps. We studied the optical and electrochemical properties of the homogeneous enzyme and also analysed the hydrolysed protein by several methods. The data presented here suggest that the carbonyl cofactor of diamine oxidase is firmly bound pyrroloquinoline quinone.
متن کاملPurification and Properties of Amine Oxidase from Epicotyls of Pisum sativum.
A procedure has been developed for the purification of amine oxidase (E.C. 1.4.3.4) from etiolated pea epicotyls (Pisum sativum cv. Little Marvel). The enzyme is sensitive to copper chelating reagents and carbonyl reagents, but is not inhibited by sulfhydryl reagents. The purified enzyme has a molecular weight of 1.85 x 10(5), as determined by sedimentation equilibrium centrifugation, and has b...
متن کاملHordeum vulgare Seedlings Amine Oxidase: Purification and Properties.
Although no amine oxidase could be detected in crude extracts, the enzyme has been purified to apparent homogeneity from Hordeum vulgare seedlings using ammonium sulfate precipitation and chromatography on DEAE cellulose, Hydroxylapatite, and Sephadex G200 columns. Gel filtration experiments indicate a molecular weight of about 150,000. The pH optimum of the enzyme was found to be 7.5 in potass...
متن کامل