Selective enrichment of environmental DNA libraries for genes encoding nonribosomal peptides and polyketides by phosphopantetheine transferase-dependent complementation of siderophore biosynthesis.
The cloning of DNA directly from environmental samples provides a means to functionally access biosynthetic gene clusters present in the genomes of the large fraction of bacteria that remains recalcitrant to growth in the laboratory. Herein, we demonstrate a method by which complementation of phosphopantetheine transferase deletion mutants can be used to restore siderophore biosynthesis and to therefore selectively enrich eDNA libraries for nonribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) gene sequences to unprecedented levels. The common use of NRPS/PKS-derived siderophores across bacterial taxa makes this method generalizable and should allow for the facile selective enrichment of NRPS/PKS-containing biosynthetic gene clusters from large environmental DNA libraries using a wide variety of phylogenetically diverse bacterial hosts.
Zachary
Z
Charlop-Powers
Jacob J
JJ
Banik
Jeremy G
JG
Owen
Jeffrey W
JW
Craig
Sean F
SF
Brady
D001483:Base Sequence; D015723:Gene Library; D005816:Genetic Complementation Test; D007501:Iron / Q000737:chemistry; D008969:Molecular Sequence Data; D015394:Molecular Structure; D046908:Peptide Biosynthesis, Nucleic Acid-Independent / Q000235:genetics*; D061065:Polyketides; D017262:Siderophores / Q000235:genetics*; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics*
D061065:Polyketides; D017262:Siderophores; D007501:Iron; D017855:Transferases (Other Substituted Phosphate Groups); C021364:holo-(acyl-carrier-protein) synthase