Purification of Mutant Forms of beta-glucosidase (His)6Zm-p60.1) Z. mays

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62156489%3A43210%2F08%3A00141770" target="_blank" >RIV/62156489:43210/08:00141770 - 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

Purification of Mutant Forms of beta-glucosidase (His)6Zm-p60.1) Z. mays

Popis výsledku v původním jazyce

beta-Glucosidases serve different roles in planta and indeed we find that plants have several different groups of this enzyme. Some of them can release active zeatin from zeatin-O-glucoside, a cytokinin conjugate thought to be the transport or storage form. One such enzyme is Zm-p60.1 from Zea mays. We have over-expressed the cDNA encoding Zm-p60.1 in E. coli and subsequently purified the recombinant protein using a purification (Zouhar et al., 1999, Filipi, 2007, Dopitova et al., 2008). It was necessary to confirm purity of wild-type, as well as mutant forms (F461L, F193A, F200K, W373K and E401D), because of its biochemical data interpretation. Unfortunately, low molecular ballast proteins were still presented in the active (dimeric) fractions of enzyme, thus the purification routine (Filipi, 2007) have been supplemented with other second gel filtration run. By addition with second gel chromatography step, enzymes yield in high homogeneity without any ballast proteins. Moreover, we ha

Název v anglickém jazyce

Purification of Mutant Forms of beta-glucosidase (His)6Zm-p60.1) Z. mays

Popis výsledku anglicky

beta-Glucosidases serve different roles in planta and indeed we find that plants have several different groups of this enzyme. Some of them can release active zeatin from zeatin-O-glucoside, a cytokinin conjugate thought to be the transport or storage form. One such enzyme is Zm-p60.1 from Zea mays. We have over-expressed the cDNA encoding Zm-p60.1 in E. coli and subsequently purified the recombinant protein using a purification (Zouhar et al., 1999, Filipi, 2007, Dopitova et al., 2008). It was necessary to confirm purity of wild-type, as well as mutant forms (F461L, F193A, F200K, W373K and E401D), because of its biochemical data interpretation. Unfortunately, low molecular ballast proteins were still presented in the active (dimeric) fractions of enzyme, thus the purification routine (Filipi, 2007) have been supplemented with other second gel filtration run. By addition with second gel chromatography step, enzymes yield in high homogeneity without any ballast proteins. Moreover, we ha

Druh

D - Stať ve sborníku

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

<a href="/cs/project/LC06034" target="_blank" >LC06034: Regulace morfogeneze rostlinných buněk a orgánů</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2008

Kód důvěrnosti údajů

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

Název statě ve sborníku

MendelNET '08 Agro

ISBN

978-80-7375-239-2

ISSN

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e-ISSN

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Počet stran výsledku

1

Strana od-do

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Název nakladatele

MZLU v Brně

Místo vydání

Brno

Místo konání akce

Brno

Datum konání akce

1. 1. 2008

Typ akce podle státní příslušnosti

EUR - Evropská akce

Kód UT WoS článku

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