The Library
Triggering the expression of a silent gene cluster from genetically intractable bacteria results in scleric acid discovery
Tools
Tools
Tools
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
|
PDF
WRAP-triggering-expression-silent-gene-cluster-bacteria-acid-Alberti-2018.pdf - Published Version - Requires a PDF viewer. Available under License Creative Commons Attribution. Download (1533Kb) | Preview |
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 | ||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Subjects: | Q Science > QD Chemistry Q Science > QR Microbiology |
||||||||||||||||||
| Divisions: | Faculty of Science > Chemistry Faculty of Science > Life Sciences (2010- ) |
||||||||||||||||||
| 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: |
|
||||||||||||||||||
| 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 | ||||||||||||||||||
| RIOXX Funder/Project Grant: |
|
||||||||||||||||||
| Open Access Version: |
Request changes or add full text files to a record
Repository staff actions (login required)
 |
View Item |
Downloads
Downloads per month over past year
