Enabling Access to Novel Bacterial Biosynthetic Potential From ONT Draft Genomic Data.

Area of Science:

• Natural Product Discovery
• Genomics
• Synthetic Biology

Background:

• Natural products offer diverse biological activities but are difficult to access.
• Soil bacteria are a rich source of novel natural products.
• Current methods limit the discovery of these valuable compounds.

Purpose of the Study:

• To develop an efficient and cost-effective pipeline for accessing biosynthetic gene clusters (BGCs) from soil bacteria.
• To leverage long-read sequencing for improved BGC identification and cloning.
• To enable the discovery of new natural products with potential therapeutic applications.

Main Methods:

• Utilized Oxford Nanopore Technology (ONT) long-read sequencing for genome assembly.
• Employed antiSMASH software for the identification of BGCs.
• Applied Transformation-associated recombination (TAR) cloning for BGC isolation.
• Tested the pipeline on the myxobacteria Aetherobacter fasciculatus SBSr002.

Main Results:

• Demonstrated the effectiveness of ONT sequencing and antiSMASH in identifying novel BGCs, even with challenges like high GC content.
• Successfully cloned a previously unknown BGC from A. fasciculatus SBSr002 into a genome engineering-ready vector.
• Validated the pipeline's capability for rapid and low-cost BGC discovery and isolation.

Conclusions:

• The developed pipeline provides a powerful and cost-effective strategy for accessing BGCs from soil bacteria.
• This approach significantly enhances the discovery potential of novel natural products.
• Facilitates the isolation of BGCs for further study and potential biotechnological applications.