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

Abstract

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.

Synergistic activity of cosecreted natural products from amoebae-associated bacteria

“…S20). These data are in accordance with previous reports that mupirocin is under QS control by 3-oxo-C10 AHL (44,45).…”

Biosynthesis of thiomarinol A and related metabolites of Pseudoalteromonas sp. SANK 73390

“…Truncated forms of mupirocin and related metabolites with C 7 and C 5 hydroxy acids have also been observed in both mutant strains and when the quorum sensing regulation mechanism has been manipulated to increase mupirocin production. 29…”

Chromosomal Arrangement of AHL-Driven Quorum Sensing Circuits in Pseudomonas

“…However, some Burkholderia species contain an additional copy of M , which is in a non-canonical arrangement, either because there are one to five additional genes betweem R and M ( M 2 topology, found in B. pseudomallei 1106a, B. pseudomallei 1710b, B. pseudomallei 668, B. pseudomallei K96243, B. thailandensis E264), or because the otherwise constitutive R gene is missing in the immediate vicinity of the MI tandem ( M 3 topology, B. ambipharia AMMD, and MC40-6). On the other hand, P. fluorescens NCIMB 10586 contains a gene coding for an enzyme, mupX in the X position [21]. …”

“…M was found to negatively regulate QS in P. fuscovaginae [24]. Finally, mupX of P. fluorescens NCIMB10586 is an amidase-hydrolase that was shown to degrade the AHL signal produced by the same species thereby decreasing the QS response [21]. …”