Mutational Analysis Reveals That All Tailoring Region Genes Are Required for Production of Polyketide Antibiotic Mupirocin by Pseudomonas fluorescens

Hothersall, Joanne; Wu, Ji En; Rahman, Ayesha; Shields, Jennifer A.; Haddock, J. W.; Johnson, Nicola; Cooper, Sian M.; Stephens, Elton R.; Cox, Russell J.; Crosby, John; Willis, Christine L.; Simpson, Thomas J.; Thomas, Christopher M.

doi:10.1074/jbc.m701490200

Cited by 38 publications

(58 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…HCS-like gene cassette has been identified in the curacin A and jamaicamide biosynthetic genes from a cyanobacterium Lyngbya majuscula [48,49], the mupirocin biosynthesis gene from Pseudomonas fluorescens, and the bacillaene biosynthesis gene from Bacillus subtilis, and is proposed to catalyze addition of C-2 from acetate onto the polyketide chain to generate a pendant functional group [50]. The pendant methyl groups in dinoflagellate polyketides might be constructed by similar genes to HCSlike gene cassette.…”

Section: Biosynthetic Studies Of Amphidinolidesmentioning

confidence: 99%

Amphidinolides and Its Related Macrolides from Marine Dinoflagellates

Kobayashi

2008

J Antibiot

144

View full text Add to dashboard Cite

This review covers the recent results described in our publications on several new cytotoxic macrolides isolated from dinoflagellates of the genus Amphidinium in addition to an overview of the isolation, structure elucidation, synthesis, biosynthesis, and bioactivity of a series of cytotoxic macrolides, named amphidinolides, reported so far. IntroductionMarine dinoflagellates, a diverse group of unicellular eukaryotes, have been recognized as real producers of marine toxins responsible to fish and algal poisoning, as well as biologically unique organisms due to their taxonomic position and unusual chromosome structure and composition. Marine dinoflagellates have also proved to be one of the most important source of bioactive natural products, which have been investigated worldwide. We have continuously studied structurally intriguing and biologically interesting bioactive macrolides and polyketides from dinoflagellates Amphidinium sp., which are symbionts of Okinawan marine flatworms Amphiscolops sp.Thirty Culture of Dinoflagellates Amphidinium sp.Large-scale cultures of the dinoflagellates of the genus Amphidinium have been performed using seawater medium enriched with Provasoli's Erd-Schriber (ES) supplement. Static incubation with illumination in a cycle of 16 hours of light and 8 hours of darkness was carried out for two weeks at 25°C. The cultures were harvested by removal of the supernatant through suction and then centrifugation to obtain algal cells. Harvested cells were extracted with MeOH -toluene, and the extracts were subjected to a systematic separation using several chromatographies to yield cytotoxic macrolides [15ϳ26]. Isolation of New Macrolides 3-1. Amphidinolactones A and BA new 13-membered macrolide, amphidinolactone A (38), C 20 H 30 O 4 , was isolated from a strain (Y-25) of a dinoflagellate Amphidinium sp. [15]. Detailed analyses of the 1 H-1 H COSY spectrum of 38 revealed connectivities of a long carbon chain from C-2 to C-20. 1 H and 13 C chemical shifts of CH 2 -2 and CH-12 suggested that C-12 was involved in an ester linkage with C-1. 1 H-1 H couplings of the two disubstituded double bonds at C-5 and C-9 indicated the Z and E geometries, respectively. Geometries of two disubstituted double bonds at C-14 and C-17 were assigned as both Z by NOESY correlations and the carbon chemical shift of C-16, which was a typical value for a methylene carbon between two Z double bonds. Thus, the gross structure of amphidinolactone A was elucidated to be 38. The relative configurations at C-8, C-11, and C-12 in 38 were deduced from NOESY correlations.A new 26-membered macrolide, amphidinolactone B (39), C 32 H 54 O 8 , has been isolated from the same dinoflagellate Amphidinium sp. (strain Y-25) as described above [16]. Detailed analyses of the 1 H-1 H COSY and TOCSY spectra as well as HMBC correlations indicated connectivities of a long carbon chain from C-2 to C-26. 1 H and 13 C chemical shifts of C-25 indicated that C-25 was involved in an ester linkage with C-1. The NOESY correlation for...

show abstract

Section: Biosynthetic Studies Of Amphidinolidesmentioning

confidence: 99%

Amphidinolides and Its Related Macrolides from Marine Dinoflagellates

Kobayashi

2008

J Antibiot

144

View full text Add to dashboard Cite

show abstract

“…Previous studies 14 failed to identify a monofunctional gene encoding a 10,11--epoxidase. We therefore consid--ered domains of multifunctional proteins.…”

Section: Analysis Of Wt and Mutant Strainsmentioning

confidence: 99%

“…Previous work on mutant strains containing single dele--tions of these genes led to the isolation of several novel PAs, including mupirocin W1 5, 15 mupirocin C2 16 14 and mupirocin F1 18a 16 (previously named mupricins W, C and F) from ∆mupW, ∆mupC and ∆mupF strains respec--tively (Figs. 3 and 4 below).…”

Section: Introductionmentioning

confidence: 99%

“…14 However, many uncertainties remain, particularly in the timing of 10,11--epoxidation, tetrahydropyran (THP) ring formation, 6--hydroxylation, fatty acid chain extension, and the pre--cise roles of, inter alia: the mupW (dioxygenase), mupC (oxidoreductase), mupF (ketoreductase), mupU (acyl CoA synthase), mupO (P450), mupV oxidoreductase), macpE (acyl carrier protein), and mmpE (ketosyn--thase/oxidoreductase) gene products.…”

Section: Introductionmentioning

confidence: 99%

“…9--HN is likely formed from a 3--hydroxy--propionate starter unit extended by three malonate condensations by MmpB functioning as an iterative FAS with additional enoyl reductase (ER) activity, perhaps provided by MupE and/or MupD. 14 The resulting backbone would need further modifications to produce the major metabolite PA--A.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Biosynthesis of Mupirocin by Pseudomonas fluorescens NCIMB 10586 Involves Parallel Pathways

Gao

Hothersall

et al. 2014

J. Am. Chem. Soc.

Self Cite

View full text Add to dashboard Cite

General rightsThis document is made available in accordance with publisher policies. Please cite only the published version using the reference above. Detailed metabolic profiling of mutant strains produced by systematic inactivation of PKS and tailoring genes, along with re--feeding of isolated metabo--lites to mutant stains, has allowed the isolation of a large number of novel metabolites, identification of the 10,11--epoxidase and the full characterisation of the mupirocin biosynthetic pathway which proceeds via major (10,11--epoxide) and minor (10,11--alkene) parallel pathways.

show abstract

Reprogramming of the Aurantinin Polyketide Assembly Line to Synthesize Auritriacids by Excising an Atypical Enoyl‐CoA Hydratase Domain

Wang,

Mao,

Zhao

et al. 2024

Advanced Science

View full text Add to dashboard Cite

Modular polyketide synthases (PKSs) are capable of synthesizing diverse natural products with fascinating bioactivities. Canonical enoyl‐CoA hydratases (ECHs) are components of the β‐branching cassette that modifies the polyketide chain by adding a β‐methyl branch. Herein, it is demonstrated that the deletion of an atypical ECHQ domain (featuring a Q280 residue) of Art21, a didomain protein contains an ECHQ domain and a thioesterase (TE) domain, reprograms the polyketide assembly line from synthesizing tetracyclic aurantinins (ARTs) to bicyclic auritriacids (ATAs) with much lower antibacterial activities. Genes encoding the ECHQ‐TE didomain proteins distribute in many PKS gene clusters from different bacteria. Significantly, the ART PKS machinery can be directed to make ARTs, ATAs, or both of them by employing appropriate ECHQ‐TE proteins, implying a great potential for using this reprogramming strategy in polyketide structure diversification.

show abstract

Mutational Analysis Reveals That All Tailoring Region Genes Are Required for Production of Polyketide Antibiotic Mupirocin by Pseudomonas fluorescens

Cited by 38 publications

References 41 publications

Amphidinolides and Its Related Macrolides from Marine Dinoflagellates

Amphidinolides and Its Related Macrolides from Marine Dinoflagellates

Biosynthesis of Mupirocin by Pseudomonas fluorescens NCIMB 10586 Involves Parallel Pathways

Reprogramming of the Aurantinin Polyketide Assembly Line to Synthesize Auritriacids by Excising an Atypical Enoyl‐CoA Hydratase Domain

Contact Info

Product

Resources

About