Shift to Pseudomonic acid B production in P. fluorescens NCIMB10586 by mutation of mupirocin tailoring genes mupO, mupU, mupV, and macpE.
Mupirocin, a polyketide-derived antibiotic from Pseudomonas fluorescens NCIMB10586, is a mixture of pseudomonic acids (PA) that target isoleucyl-tRNA synthase. The mup gene cluster encodes both type I polyketide synthases and monofunctional enzymes that should play a role during the conversion of the product of the polyketide synthase into the active antibiotic (tailoring). By in-frame deletion analysis of selected tailoring open-reading frames we show that mupQ, mupS, mupT, and mupW are essential for mupirocin production, whereas mupO, mupU, mupV, and macpE are essential for production of PA-A but not PA-B. Therefore, PA-B is not simply produced by hydroxylation of PA-A but is either a precursor of PA-A or a shunt product. In the mupW mutant, a new metabolite lacking the tetrahydropyran ring is produced, implicating mupW in oxidation of the 16-methyl group.
Cooper
Sian M
SM
School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.
Laosripaiboon
Wanpen
W
Rahman
Ayesha S
AS
Hothersall
Joanne
J
El-Sayed
A Kassem
AK
Winfield
Christopher
C
Crosby
John
J
Cox
Russell J
RJ
Simpson
Thomas J
TJ
Thomas
Christopher M
CM
D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005227:Fatty Acids / Q000096:biosynthesis / Q000235:genetics; D007533:Isoleucine-tRNA Ligase / Q000235:genetics / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D005810:Multigene Family; D016712:Mupirocin / Q000031:analogs & derivatives / Q000378:metabolism*; D009154:Mutation; D016366:Open Reading Frames; D010084:Oxidation-Reduction; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011551:Pseudomonas fluorescens / Q000235:genetics / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D005227:Fatty Acids; D048630:Polyketide Synthases; C014246:pseudomonic acid I; D016712:Mupirocin; D007533:Isoleucine-tRNA Ligase