Flonicamid (N-cyanomethyl-4-trifluoromethylnicotinamide,
FLO), a novel selective systemic pyridinecarboxamide insecticide,
effectively controls hemipterous pests. However, microbial degradation
of flonicamid, along with the enzymatic mechanism, has not been studied.
Here, bacterial isolate PG13, which converts flonicamid into 4-(trifluoromethyl)nicotinol
glycine (TFNG) and N-(4-trifluoromethylnicotinoyl)glycinamide
(TFNG-AM), was isolated and identified as Alcaligenes
faecalis CGMCC 17553. The genome of CGMCC 17553 contained
five nitrilases but no nitrile hydratase, and recombinant Escherichia coli strains harboring CGMCC 17553 nitrilase
gene nitA or nitD acquired the ability
to degrade flonicamid. Purified NitA catalyzed flonicamid into both
TFNG and TFNG-AM, indicating dual functionality, while NitD could
only produce TFNG-AM. Three-dimensional homology modeling revealed
that aromatic amino acid residues in the catalytic pocket affected
nitrilase activity. These findings further our understanding of the
enzymatic mechanism of flonicamid metabolism in the environment and
may help develop a potential bioremediation agent for the elimination
of flonicamid contamination.