title: A proteomic survey of nonribosomal peptide and polyketide biosynthesis in actinobacteria.
abstract: Actinobacteria such as streptomycetes are renowned for their ability to produce bioactive natural products including nonribosomal peptides (NRPs) and polyketides (PKs). The advent of genome sequencing has revealed an even larger genetic repertoire for secondary metabolism with most of the small molecule products of these gene clusters still unknown. Here, we employed a "protein-first" method called PrISM (Proteomic Investigation of Secondary Metabolism) to screen 26 unsequenced actinomycetes using mass spectrometry-based proteomics for the targeted detection of expressed nonribosomal peptide synthetases or polyketide synthases. Improvements to the original PrISM screening approach (Nat. Biotechnol. 2009, 27, 951-956), for example, improved de novo peptide sequencing, have enabled the discovery of 10 NRPS/PKS gene clusters from 6 strains. Taking advantage of the concurrence of biosynthetic enzymes and the secondary metabolites they generate, two natural products were associated with their previously "orphan" gene clusters. This work has demonstrated the feasibility of a proteomics-based strategy for use in screening for NRP/PK production in actinomycetes (often >8 Mbp, high GC genomes) versus the bacilli (2-4 Mbp genomes) used previously.
authors: Department of Chemistry, Northwestern University, Evanston, Illinois, 60208, United States.
authors: Yunqiu
authors: Y
authors: Chen
authors: Ioanna
authors: I
authors: Ntai
authors: Kou-San
authors: KS
authors: Ju
authors: Michelle
authors: M
authors: Unger
authors: Leonid
authors: L
authors: Zamdborg
authors: Sarah J
authors: SJ
authors: Robinson
authors: James R
authors: JR
authors: Doroghazi
authors: David P
authors: DP
authors: Labeda
authors: William W
authors: WW
authors: Metcalf
authors: Neil L
authors: NL
authors: Kelleher
mesh_terms: D039903:Actinobacteria / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis*; D005810:Multigene Family; D046908:Peptide Biosynthesis, Nucleic Acid-Independent; D010453:Peptide Synthases / Q000235:genetics / Q000378:metabolism*; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism*; D061065:Polyketides / Q000378:metabolism; D040901:Proteomics; D020539:Sequence Analysis, Protein; D053719:Tandem Mass Spectrometry
chemical_list: D001426:Bacterial Proteins; D061065:Polyketides; D048630:Polyketide Synthases; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase