About: Cyanide hydratase from Aspergillus niger K10: Overproduction in Escherichia coli, purification, characterization and use in continuous cyanide degradation     Goto   Sponge   NotDistinct   Permalink

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Description
  • A cyanide hydratase from Aspergillus niger K10 was expressed in Escherichia coli and purified. Apart from HCN, it transformed some nitriles, preferentially 2-cyanopyridine and fumaronitrile. V-max and K-m for HCN were ca. 6.8mmol min(-1) mg(-1) protein and 109mM, respectively. V-max for fumaronitrite and 2-cyanopyridine was two to three orders of magnitude lower than for HCN (ca. 18.8 and 10.3 mu.,mol min(-1) mg(-1), respectively) but K-m was also lower (ca. 14.7 and 3.7 mM, respectively). Both cyanide hydratase and nitrilase activities were abolished in truncated enzyme variants missing 18-34 C-terminal aa residues. The enzyme exhibited the highest activity at 45 degrees C and pH 8-9; it was unstable at over 35 degrees C and at below pH 5.5. The operational stability of the whole-cell catalyst was examined in continuous stirred membrane reactors with 70-mL working volume. The catalyst exhibited a half-life of 5.6 h at 28 degrees C. A reactor loaded with an excess of the catalyst was used to degrade 25 mM KCN. A conversion rate of over 80% was maintained for 3 days.
  • A cyanide hydratase from Aspergillus niger K10 was expressed in Escherichia coli and purified. Apart from HCN, it transformed some nitriles, preferentially 2-cyanopyridine and fumaronitrile. V-max and K-m for HCN were ca. 6.8mmol min(-1) mg(-1) protein and 109mM, respectively. V-max for fumaronitrite and 2-cyanopyridine was two to three orders of magnitude lower than for HCN (ca. 18.8 and 10.3 mu.,mol min(-1) mg(-1), respectively) but K-m was also lower (ca. 14.7 and 3.7 mM, respectively). Both cyanide hydratase and nitrilase activities were abolished in truncated enzyme variants missing 18-34 C-terminal aa residues. The enzyme exhibited the highest activity at 45 degrees C and pH 8-9; it was unstable at over 35 degrees C and at below pH 5.5. The operational stability of the whole-cell catalyst was examined in continuous stirred membrane reactors with 70-mL working volume. The catalyst exhibited a half-life of 5.6 h at 28 degrees C. A reactor loaded with an excess of the catalyst was used to degrade 25 mM KCN. A conversion rate of over 80% was maintained for 3 days. (en)
Title
  • Cyanide hydratase from Aspergillus niger K10: Overproduction in Escherichia coli, purification, characterization and use in continuous cyanide degradation
  • Cyanide hydratase from Aspergillus niger K10: Overproduction in Escherichia coli, purification, characterization and use in continuous cyanide degradation (en)
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  • Cyanide hydratase from Aspergillus niger K10: Overproduction in Escherichia coli, purification, characterization and use in continuous cyanide degradation
  • Cyanide hydratase from Aspergillus niger K10: Overproduction in Escherichia coli, purification, characterization and use in continuous cyanide degradation (en)
skos:notation
  • RIV/00216208:11310/14:10289017!RIV15-MSM-11310___
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • I, P(GAP504/11/0394), S
http://linked.open...iv/cisloPeriodika
  • 3
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
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  • 9489
http://linked.open...ai/riv/idVysledku
  • RIV/00216208:11310/14:10289017
http://linked.open...riv/jazykVysledku
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  • Continuous stirred membrane reactor; Aspergillus niger; Nitrilase; Cyanide hydratase (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • GB - Spojené království Velké Británie a Severního Irska
http://linked.open...ontrolniKodProRIV
  • [42907B827CB9]
http://linked.open...i/riv/nazevZdroje
  • Process Biochemistry
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http://linked.open...v/svazekPeriodika
  • 49
http://linked.open...iv/tvurceVysledku
  • Kaplan, Ondřej
  • Martínková, Ludmila
  • Plíhal, Ondřej
  • Rinágelová, Anna
  • Pasquarelli, Fabrizia
  • Veselá, Alicja Barbara
  • Cantarella, Maria
  • Chmátal, Martin
  • Křenková, Alena
http://linked.open...ain/vavai/riv/wos
  • 000334482300013
issn
  • 1359-5113
number of pages
http://bibframe.org/vocab/doi
  • 10.1016/j.procbio.2013.12.008
http://localhost/t...ganizacniJednotka
  • 11310
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