Non-Ribosomal Peptide Synthetases produce chemically diverse peptides in nature, many of which have antimicrobial properties, providing an opportunity to use synthetic biology to fine tune them for pharmaceutical applications. Major challenges remain with total and semi-synthesis of these complex peptides with specific bioengineering methodologies being developed to increase low yields and enhance bioactivity. Here we review major advances in engineering non-ribosomal peptides with a focus on improvements made to achieve better yield and bioactivity. This can be achieved through: engineering precursor metabolites, altering metabolic flux, introducing strong promoters and regulators, and redirecting metabolism to biosynthetic gene clusters which can then be expressed natively or heterologously. We also review glycopeptide antibiotics as a promising opportunity for engineering through synthetic biology for the biosynthesis of novel non-ribosomal peptides.
Keywords: Non-ribosomal peptide synthesis; antibiotics; biosynthetic gene cluster; genetic engineering; glycopeptides; heterologous expression; natural products.
The need for new antibiotics is more urgent than ever due to increasing antimicrobial resistance. This review highlights how synthetic biology has transformed the landscape of non-ribosomal peptide synthesis (NRPS) to address this crisis. By manipulating native and heterologous hosts, refactoring gene clusters, and engineering precursor and regulator elements, researchers are unlocking vast chemical diversity and improving yields of novel antibiotics. Glycopeptides are presented as a case study to showcase translational applications. This review offers a timely overview of the synthetic biology toolkit applied to NRPS and its potential to drive innovation in antibiotic discovery and development.