title: Development of an Unnatural Amino Acid Incorporation System in the Actinobacterial Natural Product Producer Streptomyces venezuelae ATCC 15439.
abstract: Many Actinobacteria, most notably Streptomyces, produce structurally diverse bioactive natural products, including ribosomally synthesized peptides, by multistep enzymatic pathways. The use of site-specific genetic incorporation of unnatural amino acids to investigate and manipulate the functions of natural product biosynthetic enzymes, enzyme complexes, and ribosomally derived peptides in these organisms would have important implications for drug discovery and development efforts. Here, we have designed, constructed, and optimized unnatural amino acid systems capable of incorporating p-iodo-l-phenylalanine and p-azido-l-phenylalanine site-specifically into proteins in the model natural product producer Streptomyces venezuelae ATCC 15439. We observed notable differences in the fidelity and efficiency of these systems between S. venezuelae and previously used hosts. Our findings serve as a foundation for using an expanded genetic code in Streptomyces to address questions related to natural product biosynthesis and mechanism of action that are relevant to drug discovery and development.
authors.affiliation: Department of Chemistry and Chemical Biology, ‡Department of Biology, and §Center for Biomedical Engineering, University of New Mexico , Albuquerque, New Mexico 87131-0001, United States.
authors.forename: Jingxuan
authors.initials: J
authors.lastname: He
authors.affiliation: Department of Chemistry and Chemical Biology, ‡Department of Biology, and §Center for Biomedical Engineering, University of New Mexico , Albuquerque, New Mexico 87131-0001, United States.
authors.forename: Briana
authors.initials: B
authors.lastname: Van Treeck
authors.affiliation: Department of Chemistry and Chemical Biology, ‡Department of Biology, and §Center for Biomedical Engineering, University of New Mexico , Albuquerque, New Mexico 87131-0001, United States.
authors.forename: Han B
authors.initials: HB
authors.lastname: Nguyen
authors.affiliation: Department of Chemistry and Chemical Biology, ‡Department of Biology, and §Center for Biomedical Engineering, University of New Mexico , Albuquerque, New Mexico 87131-0001, United States.
authors.forename: Charles E
authors.initials: CE
authors.lastname: Melançon
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000235:genetics / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D010649:Phenylalanine
keywords: Streptomyces; green fluorescent protein; natural products; p-azido-l-phenylalanine; p-iodo-l-phenylalanine; unnatural amino acids