Projects per year
Abstract
Non-ribosomal peptide biosynthesis produces highly diverse natural products through a complex cascade of enzymatic reactions that together function with high selectivity to produce bioactive peptides. The modification of non-ribosomal peptide synthetase (NRPS)-bound amino acids can introduce significant structural diversity into these peptides and has exciting potential for biosynthetic redesign. However, the control mechanisms ensuring selective modification of specific residues during NRPS biosynthesis have previously been unclear. Here, we have characterised the incorporation of the non-proteinogenic amino acid 3-chloro-β-hydroxytyrosine during glycopeptide antibiotic (GPA) biosynthesis. Our results demonstrate that the modification of this residue by trans-acting enzymes is controlled by the selectivity of the upstream condensation domain responsible for peptide synthesis. A proofreading thioesterase works together with this process to ensure that effective peptide biosynthesis proceeds even when the selectivity of key amino acid activation domains within the NRPS is low. Furthermore, the exchange of condensation domains with altered amino acid specificities allows the modification of such residues within NRPS biosynthesis to be controlled, which will doubtless prove important for reengineering of these assembly lines. Taken together, our results indicate the importance of the complex interplay of NRPS domains and trans-acting enzymes to ensure effective GPA biosynthesis, and in doing so reveals a process that is mechanistically comparable to the hydrolytic proofreading function of tRNA synthetases in ribosomal protein synthesis.
Original language | English |
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Pages (from-to) | 9466-9482 |
Number of pages | 17 |
Journal | Chemical Science |
Volume | 10 |
Issue number | 41 |
DOIs | |
Publication status | Published - 29 Aug 2019 |
Projects
- 3 Finished
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Biosynthetic LEGO: enzymatic redesign to produce new vancomycin analogues
1/01/19 → 31/12/21
Project: Research
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Development of a bio-enabled synthesis for the Glycopeptide Antibiotics
Cryle, M. & De Voss, J.
Australian Research Council (ARC), Monash University, University of Queensland
1/01/17 → 31/12/19
Project: Research
Equipment
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Monash Proteomics & Metabolomics Facility
Ralf Schittenhelm (Manager)
Faculty of Medicine Nursing and Health Sciences Research PlatformsFacility/equipment: Facility