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Triggering the expression of a silent gene cluster from genetically intractable bacteria results in scleric acid discovery
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Alberti, Fabrizio, Leng, Daniel J., Wilkening, Ina, Song, Lijiang, Tosin, Manuela and Corre, Christophe (2019) Triggering the expression of a silent gene cluster from genetically intractable bacteria results in scleric acid discovery. Chemical Science, 10 (2). pp. 453-463. doi:10.1039/C8SC03814G ISSN 2041-6520.
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Official URL: http://doi.org/10.1039/C8SC03814G
Abstract
In this study, we report the rapid characterisation of a novel microbial natural product resulting from the rational derepression of a silent gene cluster. A conserved set of five regulatory genes was used as a query to search genomic databases and identify atypical biosynthetic gene clusters (BGCs). A 20-kb BGC from the genetically intractable Streptomyces sclerotialus bacterial strain was captured using yeast-based homologous recombination and introduced into validated heterologous hosts. CRISPR/Cas9-mediated genome editing was then employed to rationally inactivate the key transcriptional repressor and trigger production of an unprecedented class of hybrid natural products exemplified by (2-(benzoyloxy)acetyl)-L-proline, named scleric acid. Subsequent rounds of CRISPR/Cas9-mediated gene deletions afforded a selection of biosynthetic gene mutant strains which led to a plausible biosynthetic pathway for scleric acid assembly. Synthetic standards of scleric acid and a key biosynthetic intermediate were also prepared to confirm the chemical structures we proposed. The assembly of scleric acid involves two unique condensation reactions catalysed by a single NRPS module and an ATP-grasp enzyme that link a proline and a benzoyl residue to each end of a rare hydroxyethyl-ACP intermediate respectively. Scleric acid was shown to exhibit moderate inhibition activity against Mycobacterium tuberculosis, as well as inhibition of the cancer-associated metabolic enzyme Nicotinamide N-methyltransferase (NNMT).
Item Type: | Journal Article | ||||||||||||||||||
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Subjects: | Q Science > QD Chemistry Q Science > QR Microbiology |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) |
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Library of Congress Subject Headings (LCSH): | Natural products -- Synthesis, Pharmaceutical microbiology, Bioactive compounds, Gene editing, Gene expression, Antibiotics -- Development | ||||||||||||||||||
Journal or Publication Title: | Chemical Science | ||||||||||||||||||
Publisher: | Royal Society of Chemistry | ||||||||||||||||||
ISSN: | 2041-6520 | ||||||||||||||||||
Official Date: | 14 January 2019 | ||||||||||||||||||
Dates: |
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Volume: | 10 | ||||||||||||||||||
Number: | 2 | ||||||||||||||||||
Page Range: | pp. 453-463 | ||||||||||||||||||
DOI: | 10.1039/C8SC03814G | ||||||||||||||||||
Status: | Peer Reviewed | ||||||||||||||||||
Publication Status: | Published | ||||||||||||||||||
Access rights to Published version: | Open Access (Creative Commons) | ||||||||||||||||||
Date of first compliant deposit: | 23 October 2018 | ||||||||||||||||||
Date of first compliant Open Access: | 23 October 2018 | ||||||||||||||||||
RIOXX Funder/Project Grant: |
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