High-level expression and characterization of Zea mays cytokinin oxidase/dehydrogenase in Yarrowia lipolytica

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15310%2F05%3A00002162" target="_blank" >RIV/61989592:15310/05:00002162 - isvavai.cz</a>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

High-level expression and characterization of Zea mays cytokinin oxidase/dehydrogenase in Yarrowia lipolytica

Original language description

Cytokinin oxidase/dehydrogenase (CKO/CKX) is a flavoenzyme, which irreversibly inactivates cytokinins by severing the isoprenoid side chain from the adenine/adenosine moiety. There are several genes coding for the enzyme in maize (Zea mays). A Z. mays CKO1 cDNA was cloned in the yeast Yarrowia lipolytica to achieve heterologous protein expression. The recombinant ZmCKO1 was recovered from cultures of transformed yeasts and purified using several chromatographic steps. The enzyme was obtained as a homogeneous protein in a remarkably high-yield and its molecular and kinetic properties were characterized. The enzyme showed a molecular mass of 69 kDa, pI was 6.3. Neutral sugar content of the molecule was 22%. Absorption and fluorescence spectra were in accordance with the presence of FAD as a cofactor. Peptide mass fingerprinting using MALDI-MS correctly assigned the enzyme in MSDB protein database. The enzyme showed a relatively high degree of thermostability (T50 = 55 °C for 30 min incub

Czech name

Vysokoúrovňová exprese a charakterizace cytokininoxidasy/dehydrogenasy ze Zea mays v Yarrowia lipolytica

Czech description

Enzym cytokinin oxidasa/dehydrogenasa (CKO/CKX) z kukuřice byl připraven jako rekombinantní protein v hostitelské kvasince Yarrowia lipolytica. Rekombinantní enzym byl izolován z kultury transformovaných kvasinek a přečištěn do homogenního stavu. Následně byly charakterizovány molekulové a kinetické vlastnosti enzymu.

Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

CE - Biochemistry

OECD FORD branch

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Project

<a href="/en/project/ME%20664" target="_blank" >ME 664: Conjugates of proteolythic enzyms in highly outputable proteomic.</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>Z - Vyzkumny zamer (s odkazem do CEZ)

Publication year

2005

Confidentiality

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

Name of the periodical

Biochimie

ISSN

0300-9084

e-ISSN

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Volume of the periodical

87

Issue of the periodical within the volume

11

Country of publishing house

FR - FRANCE

Number of pages

12

Pages from-to

1011-1022

UT code for WoS article

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EID of the result in the Scopus database

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