Genome mining for the discovery of new nitrilases in filamentous fungi.
our aim is to describe new fungal nitrilases whose sequences were published but whose catalytic properties were unknown. We adapted for expression in E. coli three of the genes and confirmed that the enzymes acted on organic nitriles. The genome mining approach was used to search for nitrilases in filamentous fungi. Synthetic genes encoding nitrilases in Aspergillus niger, Gibberella moniliformis and Neurospora crassa were expressed in Escherichia coli. This is the first heterologous expression of fungal enzymes of this type. The recombinant enzyme derived from G. moniliformis was an aromatic nitrilase with an activity of 390 U l(-1) culture with benzonitrile as substrate. This was much less than the activities of the recombinant enzymes derived from A. niger and N. crassa that had activities of 2500 and 2700 U l(-1) culture, respectively, with phenylacetonitrile as substrate.
Kaplan
Ondřej
O
Centre of Biocatalysis and Biotransformation, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20 Prague, Czech Republic.
Bezouška
Karel
K
Malandra
Anna
A
Veselá
Alicja B
AB
Petříčková
Alena
A
Felsberg
Jürgen
J
Rinágelová
Anna
A
Křen
Vladimír
V
Martínková
Ludmila
L
D000619:Aminohydrolases / Q000235:genetics / Q000378:metabolism*; D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D016681:Genome, Fungal; D005874:Gibberella / Q000201:enzymology* / Q000235:genetics; D009492:Neurospora crassa / Q000201:enzymology* / Q000235:genetics; D009570:Nitriles / Q000378:metabolism; D009930:Organic Chemicals / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D005656:Fungal Proteins; D009570:Nitriles; D009930:Organic Chemicals; D011994:Recombinant Proteins; D000619:Aminohydrolases; C021867:nitrilase