Bringing nitrilase sequences from databases to life: the search for novel substrate specificities with a focus on dinitriles

0460036 - MBÚ 2017 RIV DE eng J - Journal Article
Veselá, Alicja Barbara - Rucká, Lenka - Kaplan, Ondřej - Pelantová, Helena - Nešvera, Jan - Pátek, Miroslav - Martínková, Ludmila
Bringing nitrilase sequences from databases to life: the search for novel substrate specificities with a focus on dinitriles.
Applied Microbiology and Biotechnology. Roč. 100, č. 5 (2016), s. 2193-2202. ISSN 0175-7598. E-ISSN 1432-0614
R&D Projects: GA ČR(CZ) GAP504/11/0394; GA TA ČR TA01021368
Institutional support: RVO:61388971
Keywords : Nitrilases * Arylacetonitrilases * Dinitriles
Subject RIV: CE - Biochemistry
Impact factor: 3.420, year: 2016

The aim of this study was to discover new nitrilases with useful activities, especially towards dinitriles that are precursors of high-value cyano acids. Genes coding for putative nitrilases of different origins (fungal, plant, or bacterial) with moderate similarities to known nitrilases were selected by mining the GenBank database, synthesized artificially and expressed in Escherichia coli. The enzymes were purified, examined for their substrate specificities, and classified into subtypes (aromatic nitrilase, arylacetonitrilase, aliphatic nitrilase, cyanide hydratase) which were largely in accordance with those predicted from bioinformatic analysis. The catalytic potential of the nitrilases for dinitriles was examined with cyanophenyl acetonitriles, phenylenediacetonitriles, and fumaronitrile. The nitrilase activities and selectivities for dinitriles and the reaction products (cyano acid, cyano amide, diacid) depended on the enzyme subtype. At a preparative scale, all the examined dinitriles were hydrolyzed into cyano acids and fumaronitrile was converted to cyano amide using E. coli cells producing arylacetonitrilases and an aromatic nitrilase, respectively.
Permanent Link: http://hdl.handle.net/11104/0260185