Engineered Thiomarinol Antibiotics Active against MRSA Are Generated by Mutagenesis and Mutasynthesis of <i>Pseudoalteromonas</i> SANK73390

Murphy, Annabel C.; Fukuda, Daisuke; Song, Zhiyong; Hothersall, Joanne; Cox, Russell J.; Willis, Christine L.; Thomas, Christopher M.; Simpson, Thomas J.

doi:10.1002/anie.201007029

Cited by 39 publications

(47 citation statements)

References 20 publications

(23 reference statements)

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“…This finding is consistent with our observation regarding the substrate tolerance of HlmA, the acetyltransferase in the holomycin biosynthetic pathway. [15] The promiscuity of HolE with respect to fatty acyl CoAs suggests that it is likely responsible for the formation of the xenorhabdins ( 7 ), which were seen as pathway by-products by Thompson et al [10] …”

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confidence: 99%

Enzymatic Basis of “Hybridity” in Thiomarinol Biosynthesis

et al. 2015

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Thiomarinol is a naturally occurring double-headed antibiotic that is highly potent against methicillin-resistant Staphylococcus aureus. Its structure comprises two antimicrobial sub-components, marinolic acid and holothin, linked by an amide bond. TmlU was thought to be the sole enzyme responsible for this amide bond formation. Contrary to this idea, we show that TmlU acts as a CoA ligase that activates marinolic acid as its thioester before it is processed by the acetyltransferase HolE to catalyze the amidation. TmlU prefers complex acyl acids as substrates, whereas HolE is relatively promiscuous, accepting a range of acyl-CoA and amine substrates. Our results provide detailed biochemical information on thiomarinol biosynthesis, and evolutionary insight regarding how the marinolic acid and holothin pathways converge to generate this potent hybrid antibiotic. This work also demonstrates the potential of TmlU/HolE enzymes as engineering tools to generate new “hybrid” molecules.

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confidence: 99%

Enzymatic Basis of “Hybridity” in Thiomarinol Biosynthesis

et al. 2015

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“…23 Because the PKS and DTP pathways appeared to be independent of each other in making hybrid-antibiotic thiomarinol, mutants disrupted in PKS gene tmpD , essential for making marinolic acids, and NRPS gene holA , a homolog of holomycin biosynthetic gene hlmE , were used in a “mutasynthesis” study. 37 The Δ tmpD and Δ holA mutant bacteria were fed with alternative acids and amines respectively. The Δ tmpD strain utilized pseudomonic acid A, which differs from marinolic acid A by an extra carbon and a 4,5-oxo group, to generate a new pseudomonic acid-DTP hybrid molecule (Scheme 1a).…”

Section: Biosynthesis and Regulationmentioning

confidence: 99%

Dithiolopyrrolones: biosynthesis, synthesis, and activity of a unique class of disulfide-containing antibiotics

et al. 2014

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Dithiolopyrrolone (DTP) group antibiotics were first isolated in the early half of the 20th century, but only recently has research been reawakened by insights gained from the synthesis and biosynthesis of this structurally intriguing class of molecules. DTPs are characterized by an electronically unique bicyclic structure, which contains a compact disulfide bridge between two ene-thiols. Points of diversity within the compound class occur outside of the bicyclic core, at the two amide nitrogens. Such modifications distinguish three of the most well studied members of the class, holomycin, thiolutin, and aureothricin; the DTP core has also more recently been identified in the marine antibiotic thiomarinol, in which it is linked to a marinolic acid moiety, analog of the FDA-approved topical antibiotic Bactroban® (GlaxoSmithKline). Dithiolopyrrolones exhibit relatively broad-spectrum antibiotic activity against many Gram-positive and Gram-negative bacteria, as well as strains of Mycobacterium tuberculosis. Additionally, they have been shown to exhibit potent and selective anti-cancer activity. Despite this promising profile, there is still much unknown about the mechanisms of action for DTPs. Early reports suggested that they inhibit yeast growth at the level of transcription and that this effect is largely responsible for their distinctive microbial static properties; a similar mechanism is supported in bacteria. Elucidation of biosynthetic pathways for holomycin in Streptomyces clavuligerus and Yersinia ruckeri and thiomarinol in Alteromonas rava sp. nov. SANK 73390, have contributed evidence suggesting that multiple mechanisms may be operative in the activity of these compounds. This review will comprehensively cover the history and development of dithiolopyrrolones with particular emphasis on the biosynthesis, synthesis, biological activity and mechanism of action.

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“…Dithiolopyrrolones have been described as potent antibiotics against MRSA strains, most likely by targeting RNA polymerase 27. 30, 31 Although xenorhabdins might indeed function as antibiotics in the bacterium–nematode–insect relationship (killing other insect‐associated bacteria and thus food competitors), we thought that they might also target eukaryotic cells, as recently suggested for pyrroloformamide 32.…”

Section: Resultsmentioning

confidence: 68%