2 research outputs found
Characterization of Polyketide Synthase Machinery from the <i>pks</i> Island Facilitates Isolation of a Candidate Precolibactin
Colibactin
is a human gut bacterial genotoxin of unknown structure
that has been linked to colon cancer. The biosynthesis of this elusive
metabolite is directed by the <i>pks</i> gene cluster, which
encodes a hybrid nonribosomal peptide synthetase-polyketide synthase
(NRPS-PKS) assembly line that is hypothesized to use the unusual polyketide
building block aminomalonate. This biosynthetic pathway is thought
to initially produce an inactive intermediate (precolibactin) that
is processed to the active toxin. Here, we report the first <i>in vitro</i> biochemical characterization of the PKS components
of the <i>pks</i> enzymatic assembly line. We evaluate PKS
extender unit utilization and show that ClbG, a freestanding acyltransferase
(AT) from the <i>pks</i> gene cluster, recognizes aminomalonyl-acyl
carrier protein (AM-ACP) and transfers this building block to multiple
PKS modules, including a <i>cis</i>-AT PKS ClbI. We also
use genetics to explore the <i>in vivo</i> role of ClbG
in colibactin and precolibactin biosynthesis. Unexpectedly, production
of previously identified <i>pks</i>-associated metabolites
is dramatically increased in a Δ<i>clbP</i>/Δ<i>clbG</i> mutant strain, enabling the first structure elucidation
of a bithiazole-containing candidate precolibactin. This work provides
new insights into the unusual biosynthetic capabilities of the <i>pks</i> gene cluster, offers further support for the hypothesis
that colibactin directly damages DNA, and suggests that additional,
uncharacterized <i>pks</i>-derived metabolites containing
aminomalonate play critical roles in genotoxicity