Toggle Main Menu Toggle Search

Open Access padlockePrints

A priming cassette generates hydroxylated acyl starter units in mupirocin and thiomarinol biosynthesis

Lookup NU author(s): Professor Paul RaceORCiD


Full text for this publication is not currently held within this repository. Alternative links are provided below where available.


© 2020 American Chemical Society. Mupirocin, a commercially available antibiotic produced by Pseudomonas fluorescens NCIMB 10586, and thiomarinol, isolated from the marine bacterium Pseudoalteromonas sp. SANK 73390, both consist of a polyketide-derived monic acid homologue esterified with either 9-hydroxynonanoic acid (mupirocin, 9HN) or 8-hydroxyoctanoic acid (thiomarinol, 8HO). The mechanisms of formation of these deceptively simple 9HN and 8HO fatty acid moieties in mup and tml, respectively, remain unresolved. To define starter unit generation, the purified mupirocin proteins MupQ, MupS, and MacpD and their thiomarinol equivalents (TmlQ, TmlS and TacpD) have been expressed and shown to convert malonyl coenzyme A (CoA) and succinyl CoA to 3-hydroxypropionoyl (3-HP) or 4-hydroxybutyryl (4-HB) fatty acid starter units, respectively, via the MupQ/TmlQ catalyzed generation of an unusual bis-CoA/acyl carrier protein (ACP) thioester, followed by MupS/TmlS catalyzed reduction. Mix and match experiments show MupQ/TmlQ to be highly selective for the correct CoA. MacpD/TacpD were interchangeable but alternate trans-acting ACPs from the mupirocin pathway (MacpA/TacpA) or a heterologous ACP (BatA) were nonfunctional. MupS and TmlS selectivity was more varied, and these reductases differed in their substrate and ACP selectivity. The solution structure of MacpD determined by NMR revealed a C-terminal extension with partial helical character that has been shown to be important for maintaining high titers of mupirocin. We generated a truncated MacpD construct, MacpD_T, which lacks this C-terminal extension but retains an ability to generate 3-HP with MupS and MupQ, suggesting further downstream roles in protein-protein interactions for this region of the ACP.

Publication metadata

Author(s): Walker PD, Rowe MT, Winter AJ, Weir ANM, Akter N, Wang L, Race PR, Williams C, Song Z, Simpson TJ, Willis CL, Crump MP

Publication type: Article

Publication status: Published

Journal: ACS Chemical Biology

Year: 2020

Volume: 15

Issue: 2

Pages: 494-503

Print publication date: 21/02/2020

Online publication date: 24/01/2020

Acceptance date: 24/01/2020

ISSN (print): 1554-8929

ISSN (electronic): 1554-8937

Publisher: American Chemical Society


DOI: 10.1021/acschembio.9b00969

PubMed id: 31977176


Altmetrics provided by Altmetric