Kukuřičná beta-glukosidasa Zm-p60.1 a jejích mutantních forem: Optimalisace purifikační procedury

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62156489%3A43210%2F07%3A00117765" target="_blank" >RIV/62156489:43210/07:00117765 - isvavai.cz</a>

Výsledek na webu

—

DOI - Digital Object Identifier

—

Jazyk výsledku

angličtina

Název v původním jazyce

Maize beta-glucosidase Zm-60.1 and its mutant forms: Optimization of purification procedure

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 procedure (Zouhar et al., 1999, Verdoucq et al., 2003) which uses a phosphate buffer system, imidazole as eluent. This procedure has been successfully applied to purify the native protein, but attempts to purify His-tagged mutants were not successful. The presence of glycerol (a protection agent) in the elution solutions causesprecipitation of some mutants, and protein denaturation occurred during hydrophobic column chromatography. We therefore had to completely revise the purification protocol. For purification of recombinant enzyme and its histidine mutant f

Název v anglickém jazyce

Maize beta-glucosidase Zm-60.1 and its mutant forms: Optimization of purification procedure

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 procedure (Zouhar et al., 1999, Verdoucq et al., 2003) which uses a phosphate buffer system, imidazole as eluent. This procedure has been successfully applied to purify the native protein, but attempts to purify His-tagged mutants were not successful. The presence of glycerol (a protection agent) in the elution solutions causesprecipitation of some mutants, and protein denaturation occurred during hydrophobic column chromatography. We therefore had to completely revise the purification protocol. For purification of recombinant enzyme and its histidine mutant f

Druh

D - Stať ve sborníku

CEP obor

CE - Biochemie

OECD FORD obor

—

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

2007

Kód důvěrnosti údajů

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

Název statě ve sborníku

MendelNet'07 Agro

ISBN

978-80-7375-119-7

ISSN

—

e-ISSN

—

Počet stran výsledku

1

Strana od-do

103-103

Název nakladatele

MZLU v Brno

Místo vydání

—

Místo konání akce

Brno

Datum konání akce

1. 1. 2007

Typ akce podle státní příslušnosti

CST - Celostátní akce

Kód UT WoS článku

—