Directed natural product biosynthesis gene cluster capture and expression in the model bacterium Bacillus subtilis

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 Bacil...

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Bibliographic Details
Published in:Scientific reports 2015-03, Vol.5 (1), p.9383-9383, Article 9383
Main Authors: Li, Yongxin, Li, Zhongrui, Yamanaka, Kazuya, Xu, Ying, Zhang, Weipeng, Vlamakis, Hera, Kolter, Roberto, Moore, Bradley S., Qian, Pei-Yuan
Format: Article
Language:English
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Asparagine
Bacilli
Bacillus subtilis - genetics
Bacillus subtilis - metabolism
Bacteria
Biological Products - metabolism
Biosynthesis
Cloning
Cloning vectors
Cloning, Molecular
Escherichia coli - genetics
Gene clusters
Gene Expression Regulation, Bacterial
Genetic transformation
Genomes
Genomics
Gram-positive bacteria
Humanities and Social Sciences
Intermediates
multidisciplinary
Multigene Family
Natural products
Peptidase
Prodrugs - metabolism
Protein Biosynthesis - genetics
Recombination
Science
Surfactin
Yeasts
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Summary: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-play” 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.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep09383