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

Handle URI:
http://hdl.handle.net/10754/347333
Title:
Directed natural product biosynthesis gene cluster capture and expression in the model bacterium Bacillus subtilis
Authors:
Li, Yongxin; Li, Zhongrui; Yamanaka, Kazuya; Xu, Ying; Zhang, Weipeng; Vlamakis, Hera; Kolter, Roberto; Moore, Bradley S.; Qian, Pei-Yuan
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
Publisher:
Nature Publishing Group
Journal:
Scientific Reports
KAUST Grant Number:
SA-C0040; UK-C0016
Issue Date:
24-Mar-2015
DOI:
10.1038/srep09383
PubMed ID:
25807046
Type:
Article
ISSN:
2045-2322
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.
Additional Links:
http://www.nature.com/doifinder/10.1038/srep09383
Appears in Collections:
Publications Acknowledging KAUST Support

Full metadata record

DC FieldValue Language
dc.contributor.authorLi, Yongxinen
dc.contributor.authorLi, Zhongruien
dc.contributor.authorYamanaka, Kazuyaen
dc.contributor.authorXu, Yingen
dc.contributor.authorZhang, Weipengen
dc.contributor.authorVlamakis, Heraen
dc.contributor.authorKolter, Robertoen
dc.contributor.authorMoore, Bradley S.en
dc.contributor.authorQian, Pei-Yuanen
dc.date.accessioned2015-03-31T06:17:24Zen
dc.date.available2015-03-31T06:17:24Zen
dc.date.issued2015-03-24en
dc.identifier.citationDirected natural product biosynthesis gene cluster capture and expression in the model bacterium Bacillus subtilis 2015, 5:9383 Scientific Reportsen
dc.identifier.issn2045-2322en
dc.identifier.pmid25807046-
dc.identifier.doi10.1038/srep09383en
dc.identifier.urihttp://hdl.handle.net/10754/347333en
dc.description.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.en
dc.description.sponsorshipThis 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.en
dc.publisherNature Publishing Groupen
dc.relation.urlhttp://www.nature.com/doifinder/10.1038/srep09383en
dc.rightsThis work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/en
dc.titleDirected natural product biosynthesis gene cluster capture and expression in the model bacterium Bacillus subtilisen
dc.typeArticleen
dc.identifier.journalScientific Reportsen
dc.eprint.versionPublisher's Version/PDFen
dc.contributor.institutionSkaggs School of Pharmacy and Pharmaceutical Sciences, University of California at San Diego, La Jolla, CA 92093, United Statesen
dc.contributor.institutionDepartment of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115, United Statesen
dc.contributor.institutionJNC Corporation, Yokohama Research Center, 5-1 Okawa, Kanazawa-ku, Yokohama, Kanagawa 2368605, Japanen
dc.contributor.institutionCenter for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California at San Diego, La Jolla, CA 92093, United Statesen
dc.contributor.institutionDivision of Life Science, Hong Kong University of Science and Technology, Clear Water Bay, Hong Kongen
dc.contributor.affiliationKing Abdullah University of Science and Technology (KAUST)en
kaust.authorLi, Yongxinen
kaust.authorLi, Zhongruien
kaust.authorXu, Yingen
kaust.authorZhang, Weipengen
kaust.authorQian, Pei-Yuanen
kaust.grant.numberSA-C0040en
kaust.grant.numberUK-C0016en
kaust.grant.programKAUST Global Collaborative Research Programen
All Items in KAUST are protected by copyright, with all rights reserved, unless otherwise indicated.