1 research outputs found
Exploiting the Sensitivity of Nutrient Transporter Deletion Strains in Discovery of Natural Product Antimetabolites
Actinomycete
secondary metabolites are a renowned source of antibacterial chemical
scaffolds. Herein, we present a target-specific approach that increases
the detection of antimetabolites from natural sources by screening
actinomycete-derived extracts against nutrient transporter deletion
strains. On the basis of the growth rescue patterns of a collection
of 22 <i>Escherichia coli</i> (<i>E. coli</i>)
auxotrophic deletion strains representative of the major nutrient
biosynthetic pathways, we demonstrate that antimetabolite detection
from actinomycete-derived extracts prepared using traditional extraction
platforms is masked by nutrient supplementation. In particular, we
find poor sensitivity for the detection of antimetabolites targeting
vitamin biosynthesis. To circumvent this and as a proof of principle,
we exploit the differential activity of actinomycete extracts against <i>E. coli ΔyigM</i>, a biotin transporter deletion strain
versus wildtype <i>E. coli</i>. We achieve more than a 100-fold
increase in antimetabolite sensitivity using this method and demonstrate
a successful bioassay-guided purification of the known biotin antimetabolite,
amiclenomycin. Our findings provide a unique solution to uncover the
full potential of naturally derived antibiotics