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Statements

Subject Item
n2:RIV%2F68081707%3A_____%2F14%3A00435973%21RIV15-AV0-68081707
rdf:type
skos:Concept n17:Vysledek
dcterms:description
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. 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.
dcterms:title
Combining Rational and Random Strategies in beta-Glucosidase Zm-p60.1 Protein Library Construction Combining Rational and Random Strategies in beta-Glucosidase Zm-p60.1 Protein Library Construction
skos:prefLabel
Combining Rational and Random Strategies in beta-Glucosidase Zm-p60.1 Protein Library Construction Combining Rational and Random Strategies in beta-Glucosidase Zm-p60.1 Protein Library Construction
skos:notation
RIV/68081707:_____/14:00435973!RIV15-AV0-68081707
n3:aktivita
n11:I
n3:aktivity
I
n3:cisloPeriodika
SEP2014
n3:dodaniDat
n14:2015
n3:domaciTvurceVysledku
n9:9071105
n3:druhVysledku
n10:J
n3:duvernostUdaju
n6:S
n3:entitaPredkladatele
n5:predkladatel
n3:idSjednocenehoVysledku
7826
n3:idVysledku
RIV/68081707:_____/14:00435973
n3:jazykVysledku
n15:eng
n3:klicovaSlova
SUBSTRATE AGLYCONE SPECIFICITY; CRYSTAL-STRUCTURES; MAIZE
n3:klicoveSlovo
n13:SUBSTRATE%20AGLYCONE%20SPECIFICITY n13:CRYSTAL-STRUCTURES n13:MAIZE
n3:kodStatuVydavatele
US - Spojené státy americké
n3:kontrolniKodProRIV
[459F2A7C4955]
n3:nazevZdroje
PLoS ONE
n3:obor
n4:BO
n3:pocetDomacichTvurcuVysledku
1
n3:pocetTvurcuVysledku
4
n3:rokUplatneniVysledku
n14:2014
n3:tvurceVysledku
Brzobohatý, Břetislav Klimeš, P. Turek, D. Mazura, P.
n3:wos
000342685600060
s:issn
1932-6203
s:numberOfPages
6
n16:doi
10.1371/journal.pone.0108292