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

Abstract：

: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.

journal_name

Appl Microbiol Biotechnol

journal_title

Applied microbiology and biotechnology

authors

Veselá AB,Rucká L,Kaplan O,Pelantová H,Nešvera J,Pátek M,Martínková L

doi

10.1007/s00253-015-7023-1

subject

Has Abstract

pub_date

2016-03-01 00:00:00

pages

2193-202

issue

5

eissn

0175-7598

issn

1432-0614

pii

10.1007/s00253-015-7023-1

journal_volume

100

pub_type

杂志文章

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