TY - JOUR

T1 - Manipulation of quorum sensing regulation in Pseudomonas fluorescens NCIMB 10586 to increase mupirocin production

AU - Hothersall, Joanne

AU - Murphy, Annabel C.

AU - Iqbal, Zafar

AU - Campbell, Genevieve

AU - Stephens, Elton R.

AU - Wu, Ji'En

AU - Cooper, Helen

AU - Atkinson, Steve

AU - Williams, Paul

AU - Crosby, John

AU - Willis, Christine L.

AU - Cox, Russell J.

AU - Simpson, Thomas J.

AU - Thomas, Christopher M.

N1 - Funding information: Acknowledgement This work was supported by BBSRC grants P15257 and P07071, BBSRC/EPSRC grant E021611 employing JH, ES and AM, as well as EPSRC grant S78124 that employed JW. GC was supported by a University of Nottingham Studentship and work in PW’s laboratory was supported by BBSRC. DNA sequencing was performed by the JIF-funded Genomics Laboratory in the School of Biosciences (6/JIF13209). HPLC/LCMS equipment was funded by EPSRC grant EP/F066104). We thank Miguel Camara at the University of Nottingham for his input and comments and Ram Chhabra and Alex Truman for AHL synthesis.

PY - 2011/5/1

Y1 - 2011/5/1

N2 - Transcription of the 74 kb Pseudomonas fluorescens mupirocin [pseudomonic acid (PA)] biosynthesis cluster depends on quorum sensing-dependent regulation via the LuxI/LuxR homologues MupI/MupR. To facilitate analysis of novel PAs from pathway mutants, we investigated factors that affect mup gene expression. First, the signal produced by MupI was identified as N-(3-oxodecanoyl)homoserine lactone, but exogenous addition of this molecule did not activate mupirocin production prematurely nor did expression of mupI in trans increase metabolite production. Second, we confirmed that mupX, encoding an amidase/hydrolase that can degrade N-acylhomoserine lactones, is also required for efficient expression, consistent with its occurrence in a regulatory module linked to unrelated genes in P. fluorescens. Third, and most significantly, mupR expression in trans to wild type and mutants can increase production of antibiotic and novel intermediates up to 17-fold.

AB - Transcription of the 74 kb Pseudomonas fluorescens mupirocin [pseudomonic acid (PA)] biosynthesis cluster depends on quorum sensing-dependent regulation via the LuxI/LuxR homologues MupI/MupR. To facilitate analysis of novel PAs from pathway mutants, we investigated factors that affect mup gene expression. First, the signal produced by MupI was identified as N-(3-oxodecanoyl)homoserine lactone, but exogenous addition of this molecule did not activate mupirocin production prematurely nor did expression of mupI in trans increase metabolite production. Second, we confirmed that mupX, encoding an amidase/hydrolase that can degrade N-acylhomoserine lactones, is also required for efficient expression, consistent with its occurrence in a regulatory module linked to unrelated genes in P. fluorescens. Third, and most significantly, mupR expression in trans to wild type and mutants can increase production of antibiotic and novel intermediates up to 17-fold.

KW - Antibiotic

KW - N-acyl homoserine lactone

KW - Polyketide

KW - Quorum sensing

UR - http://www.scopus.com/inward/record.url?scp=79954724925&partnerID=8YFLogxK

U2 - 10.1007/s00253-011-3145-2

DO - 10.1007/s00253-011-3145-2

M3 - Article

C2 - 21318358

AN - SCOPUS:79954724925

VL - 90

SP - 1017

EP - 1026

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 3

ER -