. "RIV/61989592:15310/05:00002161!RIV06-MSM-15310___" . "Majira, Amelie" . "Madzak, Catherine" . . "1"^^ . "Joly, Natalie" . . "0006-3134" . "High-level expression, characterization and crystal structure of Zea mays cytokinin oxidase/dehydrogenase CKO1/CKX1." . "POLYACRYLAMIDE-GELS; POLYAMINE OXIDASE; YEAST; DEHYDROGENASE; PURIFICATION; PROTEINS; DEGRADATION; CLONING; MAIZE; ACID"@en . "9"^^ . "523306" . "Pethe, Claude" . "Houba-H\u00E9rin, Nicole" . "49" . "Vysoko\u00FArov\u0148ov\u00E1 exprese a charakterizace a krystalov\u00E1 struktura cytokinin oxidasy/dehydrogenasy CKO1/CKX1 z kuku\u0159ice (Zea mays) v Yarrowia lipolytica"@cs . "Vysoko\u00FArov\u0148ov\u00E1 exprese a charakterizace a krystalov\u00E1 struktura cytokinin oxidasy/dehydrogenasy CKO1/CKX1 z kuku\u0159ice (Zea mays) v Yarrowia lipolytica"@cs . "\u010Cl\u00E1nek je v\u011Bnov\u00E1n vysoko\u00FArov\u0148ov\u00E9 expresi, charakterizaci a krystalov\u00E9 struktu\u0159e cytokinin oxidasy/dehydrogenasy CKO1/CKX1 z kuku\u0159ice (Zea mays) v Yarrowia lipolytica"@cs . "\u0160ebela, Marek" . . "High-level expression, characterization and crystal structure of Zea mays cytokinin oxidase/dehydrogenase CKO1/CKX1."@en . . . . "[CF0D8D62331C]" . . "Cytokinin oxidase/dehydrogenase (CKO/CKX) is a flavoenzyme, which irreversibly degrades cytokinins. Zea mays CKO1 was expressed in the yeast Yarrowia lipolytica. The secreted recombinant enzyme was purified using two chromatographic steps and obtained as a homogeneous protein (12 mg/l culture). ZmCKO1 exhibited a molecular mass of 70 kD, pI of 6.3 and high thermostability (T50 = 55 oC for 30 min incubation). Neutral sugar content of the molecule was 22%. Absorption and fluorescence spectra were in accordance with the presence of FAD as a cofactor. MALDI-TOF peptide mass fingerprinting correctly assigned the enzyme in MSDB protein database. Stoichiometry of aerobic CKO reaction was assessed using spectrofluorimetry. In the presence of oxygen, ZmCKO1 produces one molecule of H2O2 per one molecule of substrate. Oxidase-mode Km values of cytokinin substrates ranged from 0.5 to 1.9 μM with pH optima varying between 7.3 and 8.0. The enzyme was crystallized in the presence of different inhibitors an"@en . "RIV/61989592:15310/05:00002161" . "Kope\u010Dn\u00FD, David" . . . . "Biologia Plantarum" . "1"^^ . "Cytokinin oxidase/dehydrogenase (CKO/CKX) is a flavoenzyme, which irreversibly degrades cytokinins. Zea mays CKO1 was expressed in the yeast Yarrowia lipolytica. The secreted recombinant enzyme was purified using two chromatographic steps and obtained as a homogeneous protein (12 mg/l culture). ZmCKO1 exhibited a molecular mass of 70 kD, pI of 6.3 and high thermostability (T50 = 55 oC for 30 min incubation). Neutral sugar content of the molecule was 22%. Absorption and fluorescence spectra were in accordance with the presence of FAD as a cofactor. MALDI-TOF peptide mass fingerprinting correctly assigned the enzyme in MSDB protein database. Stoichiometry of aerobic CKO reaction was assessed using spectrofluorimetry. In the presence of oxygen, ZmCKO1 produces one molecule of H2O2 per one molecule of substrate. Oxidase-mode Km values of cytokinin substrates ranged from 0.5 to 1.9 μM with pH optima varying between 7.3 and 8.0. The enzyme was crystallized in the presence of different inhibitors an" . "S6" . . . . "Briozzo, Pierre" . "CZ - \u010Cesk\u00E1 republika" . . "S" . . "Z(MSM6198959216)" . . "High-level expression, characterization and crystal structure of Zea mays cytokinin oxidase/dehydrogenase CKO1/CKX1." . . . "15310" . . "High-level expression, characterization and crystal structure of Zea mays cytokinin oxidase/dehydrogenase CKO1/CKX1."@en . "Laloue, Michel" . . .