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

TitleDirected natural product biosynthesis gene cluster capture and expression in the model bacterium Bacillus subtilis
Publication TypeJournal Article
Year of Publication2015
AuthorsLi Y.X, Li Z.R, Yamanaka K, Xu Y, Zhang W.P, Vlamakis H., Kolter R., Moore BS, Qian PY
JournalScientific Reports
Volume5
Date Published2015/03
Type of ArticleArticle
ISBN Number2045-2322
Accession NumberWOS:000351700300001
Keywordsabsolute-configuration; amicoumacin; bacillus-subtilis-168; cloning; competence; genome vector; heterologous expression; marfeys method; prodrug activation mechanism; protein
Abstract

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.

DOI10.1038/srep09383
Short TitleSci Rep
Student Publication: 
No
Research Topics: 
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