title: Directed natural product biosynthesis gene cluster capture and expression in the model bacterium Bacillus subtilis.
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.
authors: 1] KAUST Global Collaborative Research, Division of Life Science, Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong [2] Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California at San Diego, La Jolla, CA 92093, United States.
authors: Yongxin
authors: Y
authors: Li
authors: KAUST Global Collaborative Research, Division of Life Science, Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong.
authors: Zhongrui
authors: Z
authors: Li
authors: 1] Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California at San Diego, La Jolla, CA 92093, United States [2] JNC Corporation, Yokohama Research Center, 5-1 Okawa, Kanazawa-ku, Yokohama, Kanagawa 2368605, Japan.
authors: Kazuya
authors: K
authors: Yamanaka
authors: KAUST Global Collaborative Research, Division of Life Science, Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong.
authors: Ying
authors: Y
authors: Xu
authors: KAUST Global Collaborative Research, Division of Life Science, Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong.
authors: Weipeng
authors: W
authors: Zhang
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115, United States.
authors: Hera
authors: H
authors: Vlamakis
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115, United States.
authors: Roberto
authors: R
authors: Kolter
authors: 1] Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California at San Diego, La Jolla, CA 92093, United States [2] Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California at San Diego, La Jolla, CA 92093, United States.
authors: Bradley S
authors: BS
authors: Moore
authors: KAUST Global Collaborative Research, Division of Life Science, Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong.
authors: Pei-Yuan
authors: PY
authors: Qian
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001688:Biological Products / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D023281:Genomics; D005810:Multigene Family; D011355:Prodrugs / Q000378:metabolism; D014176:Protein Biosynthesis / Q000235:genetics*
chemical_list: D001688:Biological Products; D011355:Prodrugs