Abstract
Aspergillus niger K10 cultivated on 2-cyanopyridine produced high levels of an intracellular nitrilase, which was partially purified (18.6-fold) with a 24% yield. The N-terminal amino acid sequence of the enzyme was highly homologous with that of a putative nitrilase from Aspergillus fumigatus Af293. The enzyme was copurified with two proteins, the N-terminal amino acid sequences of which revealed high homology with those of hsp60 and an ubiquitin-conjugating enzyme. The nitrilase exhibited maximum activity (91.6 U mg-1) at 45°C and pH 8.0. Its preferred substrates, in the descending order, were 4-cyanopyridine, benzonitrile, 1,4-dicyanobenzene, thiophen-2-acetonitrile, 3-chlorobenzonitrile, 3-cyanopyridine, and 4-chlorobenzonitrile. Formation of amides as by-products was most intensive, in the descending order, for 2-cyanopyridine, 4-chlorobenzonitrile, 4-cyanopyridine, and 1,4-dicyanobenzene. The enzyme stability was markedly improved in the presence of d-sorbitol or xylitol (20% w/v each). p-Hydroxymercuribenzoate and heavy metal ions were the most powerful inhibitors of the enzyme.
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Financial support by the projects IAA4020213 (Grant Agency of the Academy of Sciences of the Czech Republic), 203/05/2267 (Czech Science Foundation), COST D25/0002/02 (European Science Foundation), LC06010 and OC D25.001 (Ministry of Education, Czech Republic), and the institutional research concept AV0Z50200510 (Institute of Microbiology) is gratefully acknowledged.
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An erratum to this article can be found online at http://dx.doi.org/10.1007/s00253-013-4743-y.
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Kaplan, O., Vejvoda, V., Plíhal, O. et al. Purification and characterization of a nitrilase from Aspergillus niger K10. Appl Microbiol Biotechnol 73, 567–575 (2006). https://doi.org/10.1007/s00253-006-0503-6
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DOI: https://doi.org/10.1007/s00253-006-0503-6