title: Recombinant thiopeptides containing noncanonical amino acids.
abstract: Thiopeptides are a subclass of ribosomally synthesized and posttranslationally modified peptides (RiPPs) with complex molecular architectures and an array of biological activities, including potent antimicrobial activity. Here we report the generation of thiopeptides containing noncanonical amino acids (ncAAs) by introducing orthogonal amber suppressor aminoacyl-tRNA synthetase/tRNA pairs into a thiocillin producer strain of Bacillus cereus .We demonstrate that thiopeptide variants containing ncAAs with bioorthogonal chemical reactivity can be further postbiosynthetically modified with biophysical probes, including fluorophores and photo-cross-linkers. This work allows the site-specific incorporation of ncAAs into thiopeptides to increase their structural diversity and probe their biological activity; similar approaches can likely be applied to other classes of RiPPs.
authors.affiliation: Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037; Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037;
authors.forename: Xiaozhou
authors.initials: X
authors.lastname: Luo
authors.affiliation: Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037; Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037;
authors.forename: Claudio
authors.initials: C
authors.lastname: Zambaldo
authors.affiliation: Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037; Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037; Department of Biology, California Institute for Biomedical Research, La Jolla, CA 92037.
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
authors.affiliation: Department of Biology, California Institute for Biomedical Research, La Jolla, CA 92037.
authors.forename: Yuhan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037; Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037;
authors.forename: Weimin
authors.initials: W
authors.lastname: Xuan
authors.affiliation: Department of Biology, California Institute for Biomedical Research, La Jolla, CA 92037.
authors.forename: Chen
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037; Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037;
authors.forename: Sean A
authors.initials: SA
authors.lastname: Reed
authors.affiliation: Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037; Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037;
authors.forename: Peng-Yu
authors.initials: PY
authors.lastname: Yang
authors.affiliation: Department of Biology, California Institute for Biomedical Research, La Jolla, CA 92037.
authors.forename: Rongsheng E
authors.initials: RE
authors.lastname: Wang
authors.affiliation: Department of Biology, California Institute for Biomedical Research, La Jolla, CA 92037.
authors.forename: Tsotne
authors.initials: T
authors.lastname: Javahishvili
authors.affiliation: Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037; Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037; Department of Biology, California Institute for Biomedical Research, La Jolla, CA 92037 schultz@scripps.edu tyoung@calibr.org.
authors.forename: Peter G
authors.initials: PG
authors.lastname: Schultz
authors.affiliation: Department of Biology, California Institute for Biomedical Research, La Jolla, CA 92037 schultz@scripps.edu tyoung@calibr.org.
authors.forename: Travis S
authors.initials: TS
authors.lastname: Young
mesh_terms: D019943:Amino Acid Substitution; D000596:Amino Acids / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001409:Bacillus cereus / Q000235:genetics / Q000378:metabolism; D015394:Molecular Structure; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D000596:Amino Acids; D010455:Peptides; D011994:Recombinant Proteins; C012574:thiocillin
keywords: antibiotic; biosynthesis; natural products; noncanonical amino acid; thiopeptides