Vysokoúrovňová exprese a charakterizace a krystalová struktura cytokinin oxidasy/dehydrogenasy CKO1/CKX1 z kukuřice (Zea mays) v Yarrowia lipolytica

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15310%2F05%3A00002161" target="_blank" >RIV/61989592:15310/05:00002161 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

High-level expression, characterization and crystal structure of Zea mays cytokinin oxidase/dehydrogenase CKO1/CKX1.

Popis výsledku v původním jazyce

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 asa 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 &#956;M with pH optima varying between 7.3 and 8.0. The enzyme was crystallized in the presence of different i

Název v anglickém jazyce

High-level expression, characterization and crystal structure of Zea mays cytokinin oxidase/dehydrogenase CKO1/CKX1.

Popis výsledku anglicky

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 asa 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 &#956;M with pH optima varying between 7.3 and 8.0. The enzyme was crystallized in the presence of different i

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

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Návaznosti

Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2005

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Biologia Plantarum

ISSN

0006-3134

e-ISSN

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Svazek periodika

49

Číslo periodika v rámci svazku

S

Stát vydavatele periodika

CZ - Česká republika

Počet stran výsledku

1

Strana od-do

"S6"

Kód UT WoS článku

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EID výsledku v databázi Scopus

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