Directed natural product biosynthesis gene cluster capture and expression in the model bacterium Bacillus subtilis
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ArticleAuthors
Li, YongxinLi, Zhongrui
Yamanaka, Kazuya
Xu, Ying
Zhang, Weipeng
Vlamakis, Hera
Kolter, Roberto
Moore, Bradley S.
Qian, Pei-Yuan

Date
2015-03-24Online Publication Date
2015-03-24Print Publication Date
2015-08Permanent link to this record
http://hdl.handle.net/10754/347333
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Bacilli 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.Citation
Directed natural product biosynthesis gene cluster capture and expression in the model bacterium Bacillus subtilis 2015, 5:9383 Scientific ReportsSponsors
This 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.Publisher
Springer NatureJournal
Scientific ReportsPubMed ID
25807046Additional Links
http://www.nature.com/doifinder/10.1038/srep09383ae974a485f413a2113503eed53cd6c53
10.1038/srep09383
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