Convergence of isoprene and polyketide biosynthetic machinery: isoprenyl-S-carrier proteins in the pksX pathway of Bacillus subtilis.
The pksX gene cluster from Bacillus subtilis is predicted to encode the biosynthesis of an as yet uncharacterized hybrid nonribosomal peptide/polyketide secondary metabolite. We used a combination of biochemical and mass spectrometric techniques to assign functional roles to the proteins AcpK, PksC, PksL, PksF, PksG, PksH, and PksI, and we conclude that they act to incorporate an acetate-derived beta-methyl branch on an acetoacetyl-S-carrier protein and ultimately generate a Delta(2)-isoprenyl-S-carrier protein. This work highlights the power of mass spectrometry to elucidate the functions of orphan biosynthetic enzymes, and it details a mechanism by which single-carbon beta-branches can be inserted into polyketide-like structures. This pathway represents a noncanonical route to the construction of prenyl units and serves as a prototype for the intersection of isoprenoid and polyketide biosynthetic manifolds in other natural product biosynthetic pathways.
Calderone
Christopher T
CT
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
Kowtoniuk
Walter E
WE
Kelleher
Neil L
NL
Walsh
Christopher T
CT
Dorrestein
Pieter C
PC
D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002070:Butadienes; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D045782:Hemiterpenes / Q000096:biosynthesis*; D008055:Lipids / Q000737:chemistry; D018942:Macrolides / Q000378:metabolism*; D013058:Mass Spectrometry; D015394:Molecular Structure; D005810:Multigene Family; D010420:Pentanes; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D002070:Butadienes; D002352:Carrier Proteins; D045782:Hemiterpenes; D008055:Lipids; D018942:Macrolides; D010420:Pentanes; C005059:isoprene; D048630:Polyketide Synthases