A Proteomic Survey of Nonribosomal Peptide and Polyketide Biosynthesis in Actinobacteria
peer-reviewed · Journal of Proteome Research · 2012
| Date | 2012-01-01 |
| Type | peer-reviewed |
| Venue | Journal of Proteome Research |
| Publisher | American Chemical Society (ACS) |
| Contribution | downstream-application |
| DOI | 10.1021/pr2009115 |
| Citations (OpenAlex) | 44 |
| Venue 2-year citedness | 3.83 |
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.
Methods and tools
- PrISM actinobacteria NRPS/PKS proteomics: Screens 26 unsequenced actinomycetes for expressed NRPS and PKS enzymes by protein-first proteomics with improved de novo sequencing, linking products to gene clusters.