caa a22 4500 397522835 CHVBK 20180308164637.0 cr unu---uuuuu 161202e199506 xx s 000 0 eng 10.1093/jxb/46.6.695 doi (NATIONALLICENCE)oxford-10.1093/jxb/46.6.695 Preliminary characterization of 1-aminocyclopropane-1-carboxylate oxidase properties from embryonic axes of chick-pea (Cicer arietinum L.) seeds [Elektronische Daten] [P. Muñoz De Rueda, M.G allardo, A.J. Matilla, I.M. Sanchez-Calle] For a deeper understanding of the germination of chick-pea (Cicer arietinum) seeds, which is dependent upon ethylene synthesis, a crude extract containing authentic ACC oxidase (ACCO) activity was isolated in soluble form from the embryonic axes of seeds germinated for 24 h. Under our optimal assay conditions (200 mM HEPES at pH 7.0, 4μM FeS04, 6 mM Na-ascorbate, 1 mM ACC, 20% 02, 3% CO2 , and 10%glycerol) this enzyme was 5-fold more active than under the conditions we used initially in the present work. The enzyme has the following Km: 28 μM for ACC (approximately 4-fold less than in vivo), 1.2% for O2 (in the presence of an optimal CO2 concentration of 3%), and 1% for CO2 in the presence of O2 (20%). The enzyme is inhibited by phenanthroline (PNT) (specific chelating agent of ferrous ion), and competitively inhibited (K1, =0.5 mM) by 2-aminoisobutyric acid (AIB), and the enzymatic activity was not detectable in the absence of CO2. Under optimal assay conditions, the enzyme has two optimum temperatures (28 °C and 35 °C) and is inhibited by divalent metal cations (Zn2+> CO2+>Ni2+>Cu2+>Mn2+ >Mg2+) and by salicylic acid, propylgallate, carbonyl cyanide m-chlorophenyl hydrazone (CCCP), dinitrophenol (DNP), and Na-benzoate. The in vitro ACCO activity which we recovered in soluble form is equivalent to approximately 80-85% of the apparent activity evaluated in vivo. © Oxford University Press Research Papers nationallicence ACC oxidase nationallicence Cicer arietinum nationallicence ethylene nationallicence germination nationallicence seeds nationallicence De Rueda P. Muñoz Departamento de Biologia Vegetal, Laboratorio Fisiologia Vegetal e Instituto de Biotecnologfa, Facultad de Ciencias, Universidad de Granada E-18001 Granada, Spain aut allardo M.G. Departamento de Biologia Vegetal, Laboratorio Fisiologia Vegetal e Instituto de Biotecnologfa, Facultad de Ciencias, Universidad de Granada E-18001 Granada, Spain aut Matilla A.J. Departamento de Biologia Vegetal, Laboratorio Fisiologia Vegetal e Instituto de Biotecnologfa, Facultad de Ciencias, Universidad de Granada E-18001 Granada, Spain aut Sanchez-Calle I.M. Departamento de Biologia Vegetal, Laboratorio Fisiologia Vegetal e Instituto de Biotecnologfa, Facultad de Ciencias, Universidad de Granada E-18001 Granada, Spain aut Journal of Experimental Botany Oxford University Press 46/6(1995-06), 695-700 0022-0957 46:6<695 1995 46 exbotj https://doi.org/10.1093/jxb/46.6.695 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/jxb/46.6.695 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- De Rueda P. Muñoz Departamento de Biologia Vegetal, Laboratorio Fisiologia Vegetal e Instituto de Biotecnologfa, Facultad de Ciencias, Universidad de Granada E-18001 Granada, Spain aut NATIONALLICENCE 700 1- allardo M.G. Departamento de Biologia Vegetal, Laboratorio Fisiologia Vegetal e Instituto de Biotecnologfa, Facultad de Ciencias, Universidad de Granada E-18001 Granada, Spain aut NATIONALLICENCE 700 1- Matilla A.J. Departamento de Biologia Vegetal, Laboratorio Fisiologia Vegetal e Instituto de Biotecnologfa, Facultad de Ciencias, Universidad de Granada E-18001 Granada, Spain aut NATIONALLICENCE 700 1- Sanchez-Calle I.M. Departamento de Biologia Vegetal, Laboratorio Fisiologia Vegetal e Instituto de Biotecnologfa, Facultad de Ciencias, Universidad de Granada E-18001 Granada, Spain aut NATIONALLICENCE 773 0- Journal of Experimental Botany Oxford University Press 46/6(1995-06), 695-700 0022-0957 46:6<695 1995 46 exbotj Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford