Combining rational and random strategies in b-glucosidase Zm-p60.1 protein library construction

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62156489%3A43210%2F14%3A00230737" target="_blank" >RIV/62156489:43210/14:00230737 - isvavai.cz</a>

Výsledek na webu

<a href="http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0108292" target="_blank" >http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0108292</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1371/journal.pone.0108292" target="_blank" >10.1371/journal.pone.0108292</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Combining rational and random strategies in b-glucosidase Zm-p60.1 protein library construction

Popis výsledku v původním jazyce

Saturation mutagenesis is a cornerstone technique in protein engineering because of its utility (in conjunction with appropriate analytical techniques) forassessing effects of varying residues at selected positions on protein structures and functions. Inthe presented study we applied the technique to randomize position W373 in b-glucosidase Zm-p60.1, whichis highly conserved among b-glucosidases. Unexpectedly, b-glucosidase activityscreening of the generated variants showed that most variants were active,although they generally had significantly lower activity than the wild type enzyme. Further characterization of the library led us to conclude that a carefully selected combination of randomized codon-based saturation mutagenesisand site-directed mutagenesis may be most efficient, particularly when constructing and investigating randomized libraries with high fractions of positive hits.

Název v anglickém jazyce

Combining rational and random strategies in b-glucosidase Zm-p60.1 protein library construction

Popis výsledku anglicky

Saturation mutagenesis is a cornerstone technique in protein engineering because of its utility (in conjunction with appropriate analytical techniques) forassessing effects of varying residues at selected positions on protein structures and functions. Inthe presented study we applied the technique to randomize position W373 in b-glucosidase Zm-p60.1, whichis highly conserved among b-glucosidases. Unexpectedly, b-glucosidase activityscreening of the generated variants showed that most variants were active,although they generally had significantly lower activity than the wild type enzyme. Further characterization of the library led us to conclude that a carefully selected combination of randomized codon-based saturation mutagenesisand site-directed mutagenesis may be most efficient, particularly when constructing and investigating randomized libraries with high fractions of positive hits.

Druh

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

CEP obor

EI - Biotechnologie a bionika

OECD FORD obor

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Projekt

<a href="/cs/project/GPP305%2F11%2FP768" target="_blank" >GPP305/11/P768: Designing proteins for understanding and modulating plant hormone metabolism and signaling</a><br>

Návaznosti

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

Rok uplatnění

2014

Kód důvěrnosti údajů

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

Název periodika

PLoS One

ISSN

1932-6203

e-ISSN

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Svazek periodika

9

Číslo periodika v rámci svazku

9

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

6

Strana od-do

1-6

Kód UT WoS článku

342685600060

EID výsledku v databázi Scopus

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