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

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    Type
    Article
    Authors
    Li, Yongxin
    Li, Zhongrui
    Yamanaka, Kazuya
    Xu, Ying
    Zhang, Weipeng
    Vlamakis, Hera
    Kolter, Roberto
    Moore, Bradley S.
    Qian, Pei-Yuan cc
    KAUST Grant Number
    SA-C0040
    UK-C0016
    Date
    2015-03-24
    Online Publication Date
    2015-03-24
    Print Publication Date
    2015-08
    Permanent link to this record
    http://hdl.handle.net/10754/347333
    
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    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-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 Reports
    Sponsors
    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 Nature
    Journal
    Scientific Reports
    DOI
    10.1038/srep09383
    PubMed ID
    25807046
    Additional Links
    http://www.nature.com/doifinder/10.1038/srep09383
    ae974a485f413a2113503eed53cd6c53
    10.1038/srep09383
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