Combining Rational and Random Strategies in beta-Glucosidase Zm-p60.1 Protein Library Construction

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F14%3A00435973" target="_blank" >RIV/68081707:_____/14:00435973 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1371/journal.pone.0108292" target="_blank" >http://dx.doi.org/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 beta-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) for assessing effects of varying residues at selected positions on proteins' structures and functions.Site-directed mutagenesis with degenerate primers is the simplest and most rapid saturation mutagenesis technique. Thus, it is highly appropriate for assessing whether or not variation at certain sites is permissible, but not necessarily the most time-and cost-effective technique for detailed assessment of variations' effects. Thus, in the presented study we applied the technique to randomize position W373 in beta-glucosidase Zm-p60.1, which is highly conserved among beta-glucosidases. Unexpectedly, beta-glucosidase activity screening of the generated variants showed that most variants were active, although they generally had significantly lower activity than the wild type enzyme.

Název v anglickém jazyce

Combining Rational and Random Strategies in beta-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) for assessing effects of varying residues at selected positions on proteins' structures and functions.Site-directed mutagenesis with degenerate primers is the simplest and most rapid saturation mutagenesis technique. Thus, it is highly appropriate for assessing whether or not variation at certain sites is permissible, but not necessarily the most time-and cost-effective technique for detailed assessment of variations' effects. Thus, in the presented study we applied the technique to randomize position W373 in beta-glucosidase Zm-p60.1, which is highly conserved among beta-glucosidases. Unexpectedly, beta-glucosidase activity screening of the generated variants showed that most variants were active, although they generally had significantly lower activity than the wild type enzyme.

Druh

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

CEP obor

BO - Biofyzika

OECD FORD obor

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Projekt

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Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

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

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Číslo periodika v rámci svazku

SEP2014

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

6

Strana od-do

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Kód UT WoS článku

000342685600060

EID výsledku v databázi Scopus

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