Enhancing armeniaspirols production through multi-level engineering of a native Streptomyces producer

• Published in: Microbial cell factories 2023-04, Vol.22 (1), p.84-8, Article 84
• Main Authors: Heng, Elena, Lim, Yi Wee, Leong, Chung Yan, Ng, Veronica W P, Ng, Siew Bee, Lim, Yee Hwee, Wong, Fong Tian
• Format: Article
• Language: English
• Subjects: Analysis; Anti-Bacterial Agents; Antibiotics; Armeniaspirols; Biological activity; Biological products; Biosynthesis; Biosynthetic engineering; Biosynthetic Pathways; Chemical properties; CRISPR; Drug resistance in microorganisms; Editing; Environmental engineering; Enzymes; Fatty acids; Fatty acyl-CoA synthase; Gene clusters; Genetic aspects; Genetic engineering; Homeopathy; Intermediates; Materia medica and therapeutics; Metabolic engineering; Metabolism; Metabolites; Methicillin; Methods; Methylation; Microbial genetic engineering; Microbiological research; Microbiological synthesis; Multigene Family; Natural products; Perturbation; Physiological aspects; Polyketides - metabolism; Production processes; Scaffolds; Staphylococcus infections; Streptomyces; Streptomyces - genetics; Streptomyces - metabolism; Therapeutics; Triglycerides; Upregulation
• ISSN: 1475-2859; 1475-2859
• DOI: 10.1186/s12934-023-02092-4

Nature has provided unique molecular scaffolds for applications including therapeutics, agriculture, and food. Due to differences in ecological environments and laboratory conditions, engineering is often necessary to uncover and utilize the chemical diversity. Although we can efficiently activate and mine these often complex 3D molecules, sufficient production of target molecules for further engineering and application remain a considerable bottleneck. An example of these bioactive scaffolds is armeniaspirols, which are potent polyketide antibiotics against gram-positive pathogens and multi-resistance gram-negative Helicobacter pylori. Here, we examine the upregulation of armeniaspirols in an alternative Streptomyces producer, Streptomyces sp. A793. Through an incidental observation of enhanced yields with the removal of a competing polyketide cluster, we observed seven-fold improvement in armeniaspirol production. To further investigate the improvement of armeniaspirol production, we examine upregulation of armeniaspirols through engineering of biosynthetic pathways and primary metabolism; including perturbation of genes in biosynthetic gene clusters and regulation of triacylglycerols pool. With either overexpression of extender unit pathway or late-stage N-methylation, or the deletion of a competing polyketide cluster, we can achieve seven-fold to forty nine-fold upregulation of armeniaspirol production. The most significant upregulation was achieved by expression of heterologous fatty acyl-CoA synthase, where we observed not only a ninety seven-fold increase in production yields compared to wild type, but also an increase in the diversity of observed armeniaspirol intermediates and analogs.