title: Escherichia coli allows efficient modular incorporation of newly isolated quinomycin biosynthetic enzyme into echinomycin biosynthetic pathway for rational design and synthesis of potent antibiotic unnatural natural product.
abstract: Natural products display impressive activities against a wide range of targets, including viruses, microbes, and tumors. However, their clinical use is hampered frequently by their scarcity and undesirable toxicity. Not only can engineering Escherichia coli for plasmid-based pharmacophore biosynthesis offer alternative means of simple and easily scalable production of valuable yet hard-to-obtain compounds, but also carries a potential for providing a straightforward and efficient means of preparing natural product analogs. The quinomycin family of nonribosomal peptides, including echinomycin, triostin A, and SW-163s, are important secondary metabolites imparting antibiotic antitumor activity via DNA bisintercalation. Previously we have shown the production of echinomycin and triostin A in E. coli using our convenient and modular plasmid system to introduce these heterologous biosynthetic pathways into E. coli. However, we have yet to develop a novel biosynthetic pathway capable of producing bioactive unnatural natural products in E. coli. Here we report an identification of a new gene cluster responsible for the biosynthesis of SW-163s that involves previously unknown biosynthesis of (+)-(1S, 2S)-norcoronamic acid and generation of aliphatic side chains of various sizes via iterative methylation of an unactivated carbon center. Substituting an echinomycin biosynthetic gene with a gene from the newly identified SW-163 biosynthetic gene cluster, we were able to rationally re-engineer the plasmid-based echinomycin biosynthetic pathway for the production of a novel bioactive compound in E. coli.
authors.affiliation: Research Core for Interdisciplinary Sciences, Okayama University, Okayama 700-8530, Japan. kenji55@cc.okayama-u.ac.jp
authors.forename: Kenji
authors.initials: K
authors.lastname: Watanabe
authors.forename: Kinya
authors.initials: K
authors.lastname: Hotta
authors.forename: Mino
authors.initials: M
authors.lastname: Nakaya
authors.forename: Alex P
authors.initials: AP
authors.lastname: Praseuth
authors.forename: Clay C C
authors.initials: CC
authors.lastname: Wang
authors.forename: Daiki
authors.initials: D
authors.lastname: Inada
authors.forename: Kosaku
authors.initials: K
authors.lastname: Takahashi
authors.forename: Eri
authors.initials: E
authors.lastname: Fukushi
authors.forename: Hiroki
authors.initials: H
authors.lastname: Oguri
authors.forename: Hideaki
authors.initials: H
authors.lastname: Oikawa
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D001412:Bacillus subtilis / Q000187:drug effects; D053898:Biosynthetic Pathways; D004448:Echinomycin / Q000031:analogs & derivatives / Q000096:biosynthesis* / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D005798:Genes, Bacterial; D005810:Multigene Family; D013302:Streptomyces / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; D004448:Echinomycin