About: Combining Rational and Random Strategies in beta-Glucosidase Zm-p60.1 Protein Library Construction     Goto   Sponge   NotDistinct   Permalink

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  • 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. (en)
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 (en)
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  • 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 (en)
skos:notation
  • RIV/68081707:_____/14:00435973!RIV15-AV0-68081707
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  • I
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  • RIV/68081707:_____/14:00435973
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  • SUBSTRATE AGLYCONE SPECIFICITY; CRYSTAL-STRUCTURES; MAIZE (en)
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  • US - Spojené státy americké
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  • PLoS ONE
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  • Klimeš, P.
  • Brzobohatý, Břetislav
  • Turek, D.
  • Mazura, P.
http://linked.open...ain/vavai/riv/wos
  • 000342685600060
issn
  • 1932-6203
number of pages
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  • 10.1371/journal.pone.0108292
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