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Nitrilase Activity Screening on Structurally Diverse Substrates: Providing Biocatalytic Tools for Organic Synthesis
Author(s) -
VergneVaxelaire Carine,
Bordier Franck,
Fossey Aurélie,
BesnardGonnet Marielle,
Debard Adrien,
Mariage Aline,
Pellouin Virginie,
Perret Alain,
Petit JeanLouis,
Stam Mark,
Salanoubat Marcel,
Weissenbach Jean,
De Berardinis Véronique,
Zaparucha Anne
Publication year - 2013
Publication title -
advanced synthesis and catalysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.541
H-Index - 155
eISSN - 1615-4169
pISSN - 1615-4150
DOI - 10.1002/adsc.201201098
Subject(s) - nitrilase , chemistry , nitrile , biocatalysis , combinatorial chemistry , high throughput screening , enzyme , carboxylic acid , metagenomics , computational biology , organic chemistry , biochemistry , catalysis , gene , reaction mechanism , biology
A high‐throughput screening of candidate nitrilases against 25 structurally diverse substrates allowed us to create a wide collection of 125 experimentally validated nitrilases. The enzymes were selected by genomic approach from 700 diverse prokaryotic species and one metagenome as representative of the nitrilase family diversity. The enzymatic screening of this collection expands the biocatalytic toolbox for chemical synthesis by providing a large number of tested nitrilases with their assigned substrates. Three examples illustrate the synthetic potential of our enzyme collection. The syntheses of carboxylic acid building blocks, a β‐substituted phenylpropanoic acid, a cyclic γ‐keto carboxylic acid and a mononitrile monocarboxylic acid, were achieved from the corresponding nitrile substrates, using three new nitrilases (two from Sphingomonas wittichii and one from Syntrophobacter fumaroxidans ). Improvements of nitrilase activities through the optimization of reaction parameters and the preparative biocatalytic synthesis are presented for these three examples.