Mild hydrolysis of nitriles by the immobilized nitrilase from Aspergillus niger K10

0039334 - MBÚ 2007 RIV SIGLE NL eng J - Journal Article
Vejvoda, Vojtěch - Kaplan, Ondřej - Bezouška, K. - Martínková, Ludmila
Mild hydrolysis of nitriles by the immobilized nitrilase from Aspergillus niger K10.
[Šetrná hydrolýza nitrilů imobilizovanou nitrilasou z Aspergillus niger K10.]
Journal of Molecular Catalysis B-Enzymatic. Roč. 39, - (2006), s. 55-58. ISSN 1381-1177
R&D Projects: GA AV ČR IAA4020213; GA ČR GA203/05/2267; GA MŠMT OC D25.001
Institutional research plan: CEZ:AV0Z50200510
Keywords : nitrilase * aspergillus niger * benzonitrile
Subject RIV: EE - Microbiology, Virology
Impact factor: 2.149, year: 2006

The cell free extract front the nitrile-hydrolyzing strain Aspergillus niger K10 (0.25 mg of protein) was adsorped onto a 1 mL HiTrap Butyl Sepharose column. The benzonitrile-hydrolyzing activity of the immobilized enzyme (about 1.6 U/mg of protein) was stable at pH 8 and 35 degrees C within the examined period (4 h). The enzyme load on the above column was increased 18 times in order to achieve high nitrile conversion. This enzyme preparation was used for the conversion of 3-cyanopyridine and 4-cyanopyridine under the above conditions. The initial substrate conversion was nearly quantitative. The activity was fairly stable; the conversion of 3-cyanopyridine decreased to 70% after 15 h, while the conversion of 4-cyanopyridine was 60% of the initial value after 39 It. The former substrate was converted into nicotinic acid and nicotinamide (molar ratio approximately 16: 1) and the latter one into isonicotinic acid and isonicotinamide (molar ratio approximately 3: 1)

Buněčný extrakt z kmene Aspergillus niger K10 hydrolyzujícího nitrily (0.25 mg proteinu) byl adsorbován na 1 ml koloně HiTrap Butyl Sepharose. Hydrolytická aktivita imobilizovaného enzymů vůči benzonitrilu (asi 1.6 U/mg proteinu) byla stabilní při pH 8 and 35 °C během 4 h. Množství vázaného enzymu bylo zvýšené 18a s cílem dosáhnout vysoké konverze nitrilu. Enzymový preparát byl použit pro konverzi 3-kyanopyridinu a 4-kyanopyridinu. Počáteční konverze substrátu byla téměř kvantitativní. Stabilita aktivity byla uspokojivá; konverze 3-kyanopyridinu se snížila na 70% po 15 h, zatímco konverze 4-kyanopyridinu byla 60% původní hodnoty po 39 h. 3-Kyanopyridin byl transformován na kyselinu nikotinovou a nikotinamid (16:1) a 4-kyanopyridin na kyselinu isonikotinovou a isonikotinamid (3:1)
Permanent Link: http://hdl.handle.net/11104/0133452