Directed natural product biosynthesis gene cluster capture and expression in the model bacterium Bacillus subtilis
Moore, Bradley S.
Permanent link to this recordhttp://hdl.handle.net/10754/347333
MetadataShow full item record
AbstractBacilli are ubiquitous low G+C environmental Gram-positive bacteria that produce a wide assortment of specialized small molecules. Although their natural product biosynthetic potential is high, robust molecular tools to support the heterologous expression of large biosynthetic gene clusters in Bacillus hosts are rare. Herein we adapt transformation-associated recombination (TAR) in yeast to design a single genomic capture and expression vector for antibiotic production in Bacillus subtilis. After validating this direct cloning plug-and-playa approach with surfactin, we genetically interrogated amicoumacin biosynthetic gene cluster from the marine isolate Bacillus subtilis 1779. Its heterologous expression allowed us to explore an unusual maturation process involving the N-acyl-asparagine pro-drug intermediates preamicoumacins, which are hydrolyzed by the asparagine-specific peptidase into the active component amicoumacin A. This work represents the first direct cloning based heterologous expression of natural products in the model organism B. subtilis and paves the way to the development of future genome mining efforts in this genus.
CitationDirected natural product biosynthesis gene cluster capture and expression in the model bacterium Bacillus subtilis 2015, 5:9383 Scientific Reports
SponsorsThis study was generously supported by a grant (DY125-15-T-02) from China Ocean Mineral Resources Research and Development Association, award SA-C0040/UK-C0016 from the King Abdullah University of Science and Technology to P.Y.Q., and funding from the NIH (R01-GM085770) to B.S.M.
PublisherNature Publishing Group
CollectionsPublications Acknowledging KAUST Support
- The logic, experimental steps, and potential of heterologous natural product biosynthesis featuring the complex antibiotic erythromycin A produced through E. coli.
- Authors: Jiang M, Zhang H, Pfeifer BA
- Issue date: 2013 Jan 13
- Clone of plipastatin biosynthetic gene cluster by transformation-associated recombination technique and high efficient expression in model organism Bacillus subtilis.
- Authors: Hu Y, Nan F, Maina SW, Guo J, Wu S, Xin Z
- Issue date: 2018 Dec 20
- Simple and rapid direct cloning and heterologous expression of natural product biosynthetic gene cluster in Bacillus subtilis via Red/ET recombineering.
- Authors: Liu Q, Shen Q, Bian X, Chen H, Fu J, Wang H, Lei P, Guo Z, Chen W, Li D, Zhang Y
- Issue date: 2016 Sep 30
- Transcriptional and functional characterization of the gene encoding acyl carrier protein in Bacillus subtilis.
- Authors: Martinez MA, de Mendoza D, Schujman GE
- Issue date: 2010 Feb
- Bacillus subtilis acyl carrier protein is encoded in a cluster of lipid biosynthesis genes.
- Authors: Morbidoni HR, de Mendoza D, Cronan JE Jr
- Issue date: 1996 Aug