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

Result code in 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>

Result on the web

<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>

Result language

angličtina

Original language name

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

Original language description

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.

Czech name

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Czech description

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Type

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

CEP classification

EI - Biotechnology and bionics

OECD FORD branch

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Project

<a href="/en/project/GPP305%2F11%2FP768" target="_blank" >GPP305/11/P768: Konstrukce proteinů pro porozmění a modulaci rostlinného hormonálního metabolismu a signalizace</a><br>

Continuities

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

Publication year

2014

Confidentiality

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

Name of the periodical

PLoS One

ISSN

1932-6203

e-ISSN

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Volume of the periodical

9

Issue of the periodical within the volume

9

Country of publishing house

US - UNITED STATES

Number of pages

6

Pages from-to

1-6

UT code for WoS article

342685600060

EID of the result in the Scopus database

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