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

Abstract

Background 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. Results 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. Conclusion 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.

Heterologous overproduction of oviedomycin by refactoring biosynthetic gene cluster and metabolic engineering of host strain Streptomyces coelicolor

“…By refactoring the ovm BGC and incorporating various engineering strategies, 670 mg/L oviedomycin was produced in a heterologous host. Similarly, Heng et al [ 53 ] applied multiple engineering strategies for the production of armeniaspirols, a class of antibiotics, by using native Streptomyces producers. These results support the need of systematic engineering strategies to significantly enhance the biosynthesis of secondary metabolites that are encoded by either silent gene clusters, or those that are produced at a low production titer.…”

Application of Cas12j for Streptomyces Editing

“…The following supporting information can be downloaded at: https: //www.mdpi.com/article/10.3390/biom14040486/s1, Figure S1: General map of all-in-one editing CRISPR-Cas construct for one-step genome editing of Streptomyces using AsCas12j-2; S1: Exconjugant outputs with the all-in-one pCRISPomyces-2 plasmids encoding for different Cas proteins. References [9,33] are cited in the supplementary materials.…”

Metabolomic analysis in Amycolatopsis keratiniphila disrupted the competing ECO0501 pathway for enhancing the accumulation of vancomycin

“…For example, the yields of amphotericin B and armeniaspirol were enhanced in S. nodosusand Streptomyces sp. A793 with the removal of the competing polyketide clusters (Huang et al 2021;Heng et al 2023). Similar observation was made previously with the improvement production of vancomycin in the mutant strainA.…”