About: Maize beta-glucosidase Zm-60.1 and its mutant forms: Optimization of purification procedure     Goto   Sponge   NotDistinct   Permalink

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  • beta-Glucosidases serve different roles in planta and indeed we find that plants have several different groups of this enzyme. Some of them can release active zeatin from zeatin-O-glucoside, a cytokinin conjugate thought to be the transport or storage form. One such enzyme is Zm-p60.1 from Zea mays. We have over-expressed the cDNA encoding Zm-p60.1 in E. coli and subsequently purified the recombinant protein using a purification procedure (Zouhar et al., 1999, Verdoucq et al., 2003) which uses a phosphate buffer system, imidazole as eluent. This procedure has been successfully applied to purify the native protein, but attempts to purify His-tagged mutants were not successful. The presence of glycerol (a protection agent) in the elution solutions causes precipitation of some mutants, and protein denaturation occurred during hydrophobic column chromatography. We therefore had to completely revise the purification protocol. For purification of recombinant enzyme and its histidine mutant forms,
  • beta-Glucosidases serve different roles in planta and indeed we find that plants have several different groups of this enzyme. Some of them can release active zeatin from zeatin-O-glucoside, a cytokinin conjugate thought to be the transport or storage form. One such enzyme is Zm-p60.1 from Zea mays. We have over-expressed the cDNA encoding Zm-p60.1 in E. coli and subsequently purified the recombinant protein using a purification procedure (Zouhar et al., 1999, Verdoucq et al., 2003) which uses a phosphate buffer system, imidazole as eluent. This procedure has been successfully applied to purify the native protein, but attempts to purify His-tagged mutants were not successful. The presence of glycerol (a protection agent) in the elution solutions causes precipitation of some mutants, and protein denaturation occurred during hydrophobic column chromatography. We therefore had to completely revise the purification protocol. For purification of recombinant enzyme and its histidine mutant forms, (en)
  • Práce řeší vylepšení purifikační rutiny isolace a purifikace rekombinantní kukuřičné beta-glukosidasy Zm-p60.1. (cs)
Title
  • Maize beta-glucosidase Zm-60.1 and its mutant forms: Optimization of purification procedure
  • Kukuřičná beta-glukosidasa Zm-p60.1 a jejích mutantních forem: Optimalisace purifikační procedury (cs)
  • Maize beta-glucosidase Zm-60.1 and its mutant forms: Optimization of purification procedure (en)
skos:prefLabel
  • Maize beta-glucosidase Zm-60.1 and its mutant forms: Optimization of purification procedure
  • Kukuřičná beta-glukosidasa Zm-p60.1 a jejích mutantních forem: Optimalisace purifikační procedury (cs)
  • Maize beta-glucosidase Zm-60.1 and its mutant forms: Optimization of purification procedure (en)
skos:notation
  • RIV/62156489:43210/07:00117765!RIV08-MSM-43210___
http://linked.open.../vavai/riv/strany
  • 103;103
http://linked.open...avai/riv/aktivita
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  • P(LC06034)
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http://linked.open...iv/duvernostUdaju
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  • 431745
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  • RIV/62156489:43210/07:00117765
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  • Zm-p60.1; optimization; purification; mutant form (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...ontrolniKodProRIV
  • [00F6401895C2]
http://linked.open...v/mistoKonaniAkce
  • Brno
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  • MendelNet'07 Agro
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  • Brzobohatý, Břetislav
  • Filipi, Tomáš
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http://linked.open.../riv/zahajeniAkce
number of pages
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  • Mendelova zemědělská a lesnická univerzita v Brně
https://schema.org/isbn
  • 978-80-7375-119-7
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  • 43210
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